Kuopion yliopiston julkaisuja A.

Farmaseuttiset tieteet 15

Kuopio University Publications A. Pharmaceutical Sciences 15

James CIaytan Callaway Jr.

Pinoline and Other Tryptamine

Derivatives: Formations and Functions

KUOPIO 1994
Kuopion yliopistonjulkaisuja A. Farmaseuttiset tieteet 15
Kuopio University Publications A. Pharmaceutical Sciences 15

James Clayton Callaway Jr.

Pinoline and Other Tryptamine

Derivatives: Formations and Functions

Doctoral dissertation

To be presented by permission of the Faculty of Pharmacy of the University

of Kuopio for public examination in Auditorium Ll, Canthia building.
University of Kuopio, on Friday_9th December 1994, at 12 noon

Department of Pharmacology and Toxicology and

Department of Pharmaceutical ChemistIy
Faculty of Pharmacy
University of Kuopio

Kuopio 1994
Distributor: Kuopio University Library
P.O.Box 1627
FIN-70211 KUOPIO
FINLAND
Tel. +35871 163430
Fax +35871 163410
,.....
,
'.
Series edlto:r: Petteri Paronen. Professor
'; , peI?~ent of Pharmaceutical Technology
University of Kuopio

Author's address: Department of Pharmaceutical Chemistry

University of Kuopio
P.O.Box 1627
FIN-70211 KUOPIO
FINLAND
Tel. +358 71 162 462
Fax +358 71 162 456
e-mail callaway@jolla.uku.fi

Supervisors: Professor Emeritus Mauno Airaksinen. M.D.

Department of Pharmacology and Toxicology
University of Kuopio

Jukka Gynther. Associate Professor

Department of Pharmaceutical Chemistry
University of Kuopio

Reviewers: Alexander T. Shulgin. Ph.D.

Department of Public Health
University of California, Berkeley, USA

Olof Beck, Associate Professor

Department of Clinical Pharmacology
Karolinska Hospital
Stockholm, Sweden

Opponents: Professor Ranan Rim6n, M.D.

Department of Clinical Psychiatry
Helsinki Central Hospital

Jyrki Taskinen, Professor

Department of Pharmaceutical Chemistry
University of Helsinki

ISBN 951-780-553-5
ISSN 1235-0478

Kuopio University Printing Office

Kuopio 1994
Finland

1994J.C. Callaway
Callaway, James C. Pinoline and other tryptamine derivatives:

formations and functions. Kuopio University Publications A.

Pharmaceutical Sciences 15 . 1994. 63 p.

ISBN 951-780-553-5

ISSN 1235-0478

ABSTRACT

The present investigations pertain to the putative formation of

tetrahydro-~-carbolines (THBCs) from serotonin (5-hydroxytryptamine, 5
HT) and other biogenic tryptamines. Some functions of these and other
tryptamine derivatives were also examined . A hypothesis was proposed
suggesting that THBCs and other indoleamines play important roles in the
regulation and maintenance of mental health . Earlier hypotheses have
considered only the dysfunctional aspects of the 'aberrant' formation of
endogenous psychoactive agents . The literature on endogenous
tryptamine derivatives is reviewed, and functional aspects of such agents
are discussed .
I n vitro NMR spectroscopic experiments demonstrated that I-H- THBCs
(THBCs with only hydrogen atoms at position I) may be formed from
biogenic tryptamines and glyoxylic acid, though not from formaldehyde at
physiologic pH, and that I-Me-THBCs may be formed from acetaldehyde,
though not pyruvic acid, under these conditions. These results challenge
previous declarations on the formation of THBCs, and verify earlier
observations .
Pinoline, an endogenous THBC with demonstrated serotonergic actions,
was tritiated and receptor binding characteristics were investigated.
[3 H)Pinoline was found to have good affinity for the 5-HT uptake site when
compared with specific 5-HT uptake inhibitors such as paroxetine and
citalopram . Receptor binding and auto radiographic studies add support to
the suggestion that biological pinoline may function as a natural ligand
for the 5-HT uptake site, which has relevance to affective disorders. In
addition to the successful synthesis and characterization of [3 HJ pinoline, a
method was described to determine the specific activity of a tritiated
species by NMR.
Long-term effects from the periodic ingestion of aya h uasca, typically a
beverage of N,N-dimethyltryptamine (DMT) and ~-carbolines, were
associated with an increased density of 5-HT uptake sites in human blood
platelets, when compared with controls. This observation was novel and
may also be of significance in the treatment of affective disorders.

National Library of Medicine Classification: QV 126, QV 77.5, QV 77.7. QV 38,

WL 104

Medicinal Subject Headings: indoles; carbolines; tryptamines; serotonin;

receptors. serotonin; neurotransmitter uptake inhibitors; N,N
dimethyltryptamine; harmaline; harmine; monoamine oxidase inhibitors;
biogenic amines; citalopram; pineal body; blood platelets; psychotropic
drugs; human; medicine. traditional; affective disorders/drug therapy;
depression/drug therapy; autoradiography; nuclear magnetic resonance
'Insanity is the dream of the man who is awake'

J.1. Moreau de Tours, 1804-1884

To Anita
ACKNOWLEDGEMENTS

The present work was carried out in the Department of PharTnaceutical

Chemistry and in the Department of Pharmacology and Toxicology,
University of Kuopio, Finland, during the years 1990-1994.
It is my pleasure to gratefully acknowledge the continuing advice,
support and inspiration I've received from Professor Emeritus Mauno
Airaksinen M.D., Ph.D. (Pharmacology), the discoverer of pinoline. The
present work would not have succeeded without his supervision and vast
experience in the area of neuroactive tryptamine derivatives. Hopefully,
his recent retirement will allow him even more time to pursue his
interests in this fascinating field .
also wish to express my deepest appreciation to Associate Professor
lukka Gynther, Ph.D. (Pharm.) for his brotherly guidance. valuable advice
and encouragement. He was never too busy to help me with even the most
obvious problems. I also thank Pekka Peura. Ph .D. (Pharm) for his advice,
in addition to his kind and considerate interest in this work.
I also thank Alexander T. Shulgin. Ph.D. (Org. Chem.) and Associate
Professor Olof Beck, Ph.D. (Pharmacol.) for agreeing to review this
manuscript. Their contributions to this work were highly appreciated.
I am likewise beholden to Dave Pate, M.Sc. (Bio.). for my introduction to
the ~-carbolines and thanks to 10 Falcon B.A. (Anthro.) for proofreading
this manuscript. I would also like to thank Peyton lacob, Ph.D. (Pharm.
Chem.) for his mechanistic insights.
Also I thank those who have directly participated in the production of
this work; Glacus Brito. M.D., Charles Grob, M.D. (Psych.), Eija Kari, M.Sc.
(Pharm.), Dennis McKenna, Ph.D. (Bot.), Hiromi Morimoto. M.Sc. (Chem.),
Petri Nykvist, M.Sc. (Pharm.), Antti Poso, M.Sc. (Pharm.), Lembit Rligo,
M.D., Ph.D. (Pharmacol.), 10uko Veps!tl!tinen, Ph.D. (Chem.), Philip
Williams, Ph.D. (Chem).
In addition, my sincere thanks go out to all my friends, colleagues and
staff within the School of Pharmacy at University of Kuopio with whom
I've had the pleasure to know during this time. I especially thank the
following for many things: Seppo Auriola, Ph.D. (Pharm.), Mr. luhani
Hanninen, Tomi llirvinen, Ph .D. (Pharm .), Mr. lukka Knuutinen, Kirsti
Laitinen M.Sc. (Pharm.), Leena Linqvist, Ph.D. (Biochem.). Ewen Mac
Donald. Ph.D. (Pharm.). Jukka Monkkonen, Ph.D. (Pharm.), Toivo
Nauranlahti, Ph .D. (Pharm .). Mrs. Kaarina Pitklinen. Cecil Navajas Polo,
Lic. (Pharm .). Veli-Pekka Ranta, M.Sc . (Pharm.), Olavi Raatikainen, Lic.
(Phi!.) . Pekka Suhonen, M.Sc. (Pharm .). Hannu Taipale. M.Sc. (Pharm .),
Tuula Tennilli, M.sc. (Pharm.) and Mrs. Eila Turunen .
I also thank Jukka Hiltunen, Managing Director of MAP Medical
Technologies Oy. for the connection to Phil Williams at the National
Tritium Labeling Facility (NTLF). Lawrence Berkeley Laboratories (LBL),
Berkeley, California.
Many thanks to Drs. Victor Py-Danieal and Jose Cabral of the Instituto
Nacional De Pesquisa Da Amazonia (INPA) in Manaus. Brazil, for the use of
their facilities, and to the members of the Uniao Do Vegetal for their
gracious invitation to study the Vegetal.
I also thank the University of Kuopio for annual travel support to
international conferences.
Finally. I would like to express my deepest gratitude to Anita Hemmila.
for her enduring patience and for her interest in these explorations. and
also for allowing our home life to revolve around the effects of ~

carbolines and other tryptamine derivatives for the past 12 years.

This work has been funded in part from the Medical Research Council.
Academy of Finland, the Finnish Cultural Fund. and Botanical Dimensions
(USA), a non-profit research organization supporting tbe investigation of
ethnomedically significant plants .

Kuopio, November 1994

Year of the Dog

)t(
Jace Callaway
ABBREVIATIONS

~ beta
BCs ~-carbolines
CH3 methyl (functional group, also as H3C)
CNS central nervous system
COOH carboxylic acid (functional group)
CSF cerebrospinal fluid
m-IBC 3 ,4-d i hydro - ~-carboline
DHBCs 3 ,4-di hydro-~-carbol ines
DMf N .N -d imethyl tryptamine
H hydrogen (atom)
H3C methyl (functional group, also as CH3)
H3CO methoxy (functional group, also as MeO)
HIOMT 5 -hydroxyindole-O -methyltran sferase
HO hydroxy (functional group)
5-HODMT 5-hydroxy-DMT, bufotenine
5-HT 5-hydroxytryptamine, serotonin
I-H-THBC l,2,3,4-tetrahydro-~-carboline, THBC
I-H-THBCs 1,2,3 ,4-tetrahydro-~-carbolines, THBCs
MAO monoamine oxidase (enzyme)
MAO-A monoamine oxidase type A (enzyme)
Me methyl (also written as CH3 or H3C)
MeO methoxy (also written as CH30 or OCH3)
5-MeODMT 5 -methox yd imethyl tryptami ne
6-MeO-DHBC 6-methoxy-3,4-DHBC, didehydro-pinoline
5-MeO-N-MeT 5 -methoxy-N -methyltryptamine
5-MeOT 5 -methoxytryptamine
6-MeO-THBC 6-methoxy-tetrahydro- ~ - carboline (pinoline)
5 - MeO-tryptoline 6- MeO-THBC, pinoline
N - MeT N -methyltryptamine
I-Me-THBC I-meth yl-tetrahydro- ~-carbol ine
I-Me-THBCs I-methy I-tetrahydro- ~-c arboli nes
NAT N -acetyltransferase
NMT N -methy Itransferase
NMR nuclear magnetic resonance (spectroscopy)
PEA phenethylamine
R general symbol for varying substituents
T tryptamine
THBC 1,2,3 ,4-tetrahydro-~-carboline, I-H-THBC
THBCs tetrahydro-~-carbolines, I-H-THBCs
LIST OF ORIGINAL PUBLICATIONS

This work is based on the following papers referred to in the text

by Roman numerals I-VI:

I Callaway l.C.: A proposed mechanism for the visions of

dream sleep. Medical Hypotheses 26: 119-124, 1988

I I Callaway l.C., Morimoto H., Gynther l., Airaksinen M.M. and

Williams P.G.: Synthesis of [3H]pinoline, an endogenous
tetrahydro-~-carboline. 1 Labelled Compds Radiopharm 31:

355-364, 1991

I I I Airaksinen M.M., Callaway l.C., Nyqvist P., Rago L., Kari E.

and Gynther l.: Binding sites for [3H]pinoline, pp. 83-86. In:
Melatonin and the Pineal Gland: From Basic Science to
Clinical Application, Touitou Y, Arendt land Pevet P, eds.
International Congress Series 1017, 1993. Elsevier Science
Publications: Amsterdam

I V Callaway l.C., Gynther l., Poso A., Vepsalainen l. and

Airaksinen M.M.: The Pictet-Spengler reaction and biogenic
tryptamines: formation of tetrahydro-~-carbolines at
physiological pH. 1 Heterocyclic Chem 31 (2):431-435, 1994

V Callaway l.C., Airaksinen M.M. and Gynther l.: Endogenous

~-carbolines and other indole alkaloids in mammals.
Integration 5:1-14, 1994

V I Callaway l.C., Airaksinen M.M., McKenna ~J., Brito G.S ., Grob

C.S.: Platelet serotonin uptake sites increased in drinkers of
ayahuasca. Psychopharmacology (in press), 1994
CONTENTS

1 INTRODUCTION 17

2 AIMS OF THE STUDY 28

3 MATERIALS AND METHODS 29

3.1 Chemicals 29
3.2 Synthesis of the ~-carbolines 29
3.2.1 Synthesis of THBCs 29
3.2.2 Synthesis of DHBCs 29
3.2.3 Synthesis of [3H]pinoline 30
3.3 Characterization of the products 30
3.3.1 Thin layer chromatography 30
3.3 .2 Mass spectrometric analysis 30
3.3.3 Nuclear magnetic resonance (NMR) analyses 3 1
3.3.3.1 IH and 13C-NMR analyses 31
3.3.3.2 IHf3H-NMR analyses 31
3.3.3.3 IH-NMR reaction kinetics 32
3.3.4 MOPAC analysis of reaction intermediates 32
3.4 Binding studies of the 5-HT uptake site 33
3.4.1 [3H]Citalopram 33
3.4.2 [3H]Pinoline 33
3.4.3 Analysis of the binding data ' 33
3.5 Autoradiography with [3H]pinoline 34

4 RESULTS 35
4.1 Formation of THBCs 35
4.1.1 Endogenous formation of THBCs 35
4.1.2 Synthesis and characterization of [3H]pinoline 36
4.2 Some functions of THBCs and other indoleamines 37
4.2.1 Binding characteristics of [3H]pinoline 37
4.2.2 Putative functions of endogenous indoles 38
4.2.3 Some effects of exogenous indoles 38
5 DISCUSSION 39
5.1 Fonnations of THBCs 39
5.1.1 In vitro formation from aldehydes 39
5.1.1.1 Fonnation of 1-H -THBCs from formaldehyde 40
5.1.1.2 Formation of 1-Me-THBCs from acetaldehyde 41
5.1.2 In vitro formation from a-keto acids 4I
5.1.2.1 Formation of 1-H-THBCs from glyoxylic acid 41
5.1.2.2 Fonnation of 1-Me-THBCs from pyruvic acid 42
5.1.3 Biological formations 42
5.1.4 Synthesis and characterization of [3H]pinoline 43
5.1.4.1 Synthesis of [3H]pinoline 43
5.1.4.2 Characterization of [3H]pinoline 44
5.2 Some functions of THBCs 45
5.2.1 Binding to the 5-HT uptake site 45
5.2.2 Psychoactivity of THBCs and other BCs 46
5.2.3 Putative functions of endogenous THBCs 47
5.2.4 Putative dysfunctions of endogenous THBCs 48
5.3 Exogenous indoles and the 5-HT uptake site 49

6 CONCLUSIONS 52

7 REFERENCES 54

ORIGINAL PUBLICATIONS 63
 17

1 INTRODUCTION

A vast array of neurochemical mechanisms is reasoned to process

sensory information and facilitate the expression of behavior (Eccles
1992), yet absolute correlates linking neuroact!vlty with
psychoactivity remain obscure. The investigations described herein
focus on a narrow aspect of this multifaceted system; specifically
the phenomena associated with the neurotransmitter serotonin (5
hydroxytryptamine, 5-HT) and other tryptamine derivatives (Fig.
I). Such compounds are also known collectively as indoleamines, or
sometimes as indolealkylamines.
Drugs that precipitate changes in mental activity have been found
to modify serotonergic activity within the central nervous system
(CNS). Thus, some anomalies in behavior are treated with drugs that
have affinity for 5-HT receptor sites.
Psychedelic drugs, literally 'mind-revealing' drugs, also have
demonstrated actions at 5-HT sites in pharmacological tests and
allow reliable access to extraordinary states of mind. These unique
compounds have also been known as psychotropic, hallucinogenic,
psychotomimetic, psychotogenic, and entheogenic drugs. (A brief
discussion of terminology has been recently published; Call away
1993 ).
The tryptamines (1H -indole-3-ethanamines) are a class of
indoleamines that effect serotonergic systems (Airaksinen and
McIsaac 1968, Ho et al. 1970). Tryptamines such as 5-HT and N ,N
dimethyltryptamine (DMT) are characterized by an aromatic
benzopyrene more commonly known as the indole (Fig. I). While
some of the indoleamines discussed herein are in fact tryptamines,
others more central to these investigations belong to a related class
of compounds known as the p-carbolines (9H -pyrido[3 ,4-b]indoles;
Fig. 2).
 I8

Indole Tryptamines

Tryptamines Abbr. RI R2

tryptami ne T H H
serotonin 5-HT HO H
5 -methoxytryptamine 5-MeOT MeO H
N ,N -dimethyltryptamine DMT H CH3
bufotenine 5-HODMT HO CH3
5-methoxy-DMT 5-MeODMT MeO CH3

Fig.I. The chemical indole, its numbering system, and some

tryptamine derivatives.

6 5 4 3

7Q-CN2
8 N 1

OcCN N
OcCNH N
H H H

~-carboline d ihydro- ~-carboline tetrahydro- ~-carboline

Fig. 2. ~-carboline (BC) and numbering system, 3,4-dihydro-~

carboline (DHBC) and I ,2,3,4-tetrahydro-~-carboline (THBC).
 19

The prevailing wisdom of the scientific literature on ~-carbolines

(BCs) holds that these indoleamines are capable of producing
hallucinations in humans (Pennes and Hoch 1957, N ararijo 1967).
Contrary to earlier reports, however, the BCs do not readily bring
about hallucinogenic or even psychedelic activity on their own
(Shulgin 1977, Call away 1993, Ott 1994). This confusion dates back
to a time when a subset of BCs, known collectively as harmala
alkaloids (Fig. 3), were thought to be the only psychoacti ve
alkaloids in the sacred Amazonian beverage ayahuasca. This 'tea' is
known by many names throughout the region, the name ayahuasca
now being most prevalent in contemporary literature (McKenna and
Towers 1984). Ayahuasca is typically prepared from Banisteriopsis
c a a pi, a jungle vine rich in harmine, harmaline and
tetrahydroharmine. DMT (Fig.l), another typical component of
ayahuasca, is obtained from the leaves of Psychotria viridis.

harmine harmaline tetrahydroharmine

Fig. 3. The major harmala alkaloids found in ayahuasca.

Excellent descriptions of both the cultural and

psychopharmacological signi ficance of ayahuasca have been
pu blished (Luna and Amaringo 1991 and Ott 1994, respectively).
Richard Spruce may have been the first academician to seriously
study the tea, known to him in 1851 only as caapi (Spruce 1873),
though earlier reports from enthusiastic Jesuit missionaries had
referred to such beverages as 'diabolical brews' (Ott 1994). The tea
 20

is also known as yaje, hoasca, vegetal, daime, and natem, to name a

few.
The primary function of the harmala alkaloids in ay a h u a sea
(harmine in particular) is to allow for the oral activity of DMT by
inhibition of the enzyme monoamine oxidase type A (MAO-A). DMT
is a powerful psychedelic agent that is quickly metabolized, and
otherwise inactive upon oral administration (Holmstedt & Lindgren
1967, Shulgin 1976). Profound visual imagery floods the mind for
only a few minutes when DMT is smoked or injected (Strassman et
at. 1994).
The inhibition of MAO-A to facilitate the efficacy of an otherwise
orally inactive substance (DMT) distinguishes ayahuasca from other
pharmacological preparations. Enzymatic inhibition not only allows
for the oral activity of DMT, but also the accumulation of 5-HT and
other neurotransmitters which are ordinarily metabolized by this
enzyme. Because of this complex chemical synergy, there has been
much confusion in the scientific literature over the effects of
ayahuasca and its relationship to the psychoactivity of BCs in
general.
One of the earliest mentions of pharmacologic activity from BCs,
as plant derived harmala alkaloids, can be found in the Greek
Herbal of Dioscorides (De Materia Medica; a leading pharmacological
text for 16 centuries). This important text was compiled in 77 AD
and describes the effects of pulverized seeds from Peg an u m
harmala as 'being good for the dullness of sight'. Other than being
'extremely bitter to the taste', no other information of immediate
pharmacological interest was offered (Gunter 1934).
Another early reference to P. harmala can be found in a Mandean
scroll from the 9th century, believed to have been translated from
an earlier text, entitled 'Safta d Sambra' (The Scroll of [Wild] Rue),
v
where the plant is quoted a stating 'I am Sambra, the good
neighbor, king of all drugs' (Drower 1934). The Zoroastrians are
another religious group of the Indus Valley who may have
employed a preparation of P. harmala as the sacramental beverage
Ha 0 m a, and recent speculation has attempted to link this plant
 21

teacher and its chemical constituents to the Soma of ancient Vedic

hymns (Flattery and Schwartz 1989). Since reasonable doses of the
harmala alkaloids lack significant psychoactivity on their own, it
seems unlikely that P. harmala could have been solely responsible
for the religious effects of this mythical beverage. However, in
combination with a local plant source of DMT, such as Desmodium or
Phalaris species, a beverage analogous to ayahuasca could certainly
have produced an efficacious sacrament worthy of Vedic praise!
In the first known compendium of psychoactive plants, Olof
Alander (1762), under the direction of Karl Linnaeus, attributes the
following psychoactive properties to the seeds of P. harmala :

PEGANUM Harmala, its seeds are taken as an inebriant by

the Turks, the writer Bellonio reports in his itinerary [with
God as his witness], that the Emperor Solimannum was
accustomed to eating these seeds, but the author did not
know what kind of substance could provoke such delight
or erase recollection of disagreeable things. Evidently it
was a dish made of the seeds that Krempherus ate at a
gathering during his travels in Persia, which caused him to
be overcome with inexpressible joy such as he had never
previously experienced, with caresses, laughter, hilarity,
etc., at the end of the meal, when he mounted his horse,
his brain was imp1anted with an extraordinary sense of
power, he felt virtually as if he were astride Pegasus and
was flying through the clouds and along multicolored
rainbows in the sky, and even as if he was having dinner
with the gods; by the following day he had completely
forgotten what had happened. (DuQuesne 1983)

However, neither of Alander's two sources had bothered to identify

these mysterious seeds. From the effects described by the German
born physician/traveler Krempherus (Kaempfer, Engelbert 1651
 22

1716) it seems more likely that a dish of seeds other than P.

harmala had actually been consumed.
For a contemporary account of the psychoactive effects of
harmala alkaloids, the following reference by Ott is recommended
(Ott 1994). Although having only weak psychoactive effects, the
harmala alkaloids do possess enough neuroactivity to have
interested ancient and modern psychopharmacologists over the
years; i.e. as inhibitors of MAO-A and thus subsequent potentiators
of monoaminergic activity in the CNS (Buckholtz and Boggan 1977).
In animals, tryptamines and BCs can enter the body directly
through the diet or, for the most part, be produced endogenously
from dietary tryptophan. In the biological production of 5-HT, for
example, the essential amino acid L-tryptophan is converted to 5
hydroxy tryptophan by the enzyme tryptophan hydroxylase, then
decarboxylated to 5-HT. This neurotransmitter (5-HT) was first
crystalized from mammalian blood in 1948, and named se roto ni n
since 'its source is serum and its activity is one of causing
constriction' (Rapport et al. 1948). Other tryptamines are also
produced from dietary tryptophan (Fig. 4).
1,2,3,4- Tetrahydro-~ -carbolines (THB Cs) are readily produced
from biogenic tryptamines through a Pictet-Spengler condensation
with an appropriate carbonyl source under laboratory or
physiological conditions (Fig. 5). This reaction was originally
described for the formation of tetrahydroisoquinolines from
phenethylamines (Picfet & Spengler 1911) and subsequently
applied to the formation of THBCs from tryptamines.
The search for endogenous THBCs, and other tryptamine
derivatives, as biochemical explanations for mental illness began in
the early 1960's as an extension of the transmethylation hypothesis
for schizophrenia (Osmond and Smythies 1952). In 1961, McIsaac
reported the identification of 'adrenoglomerulotropin' (6-methoxy
1-methyl-tetrahydro-~-carboline, 6-MeO-1-Me-THBC,) in the urine

of rats after treatment with 5-methoxytryptamine, ethanol,

disulfiram (Antabuse), and iproniazid (McIsaac 1961). The logic in
this experiment was straightforward. From ethanol metabolism,
 23

o
~ ~OH
'-0 NH,
N
H ~YdrOXYI:tion

L-tryptophan
NH,
HO~
~
-- OH
~ A ~ NH,
N N
H H

tryptamine 5 -hydroxy tryptop han

!
I NMT

NHCH,
HO

~'
Decarboxylation

NH

~ ~
~ N
NMT / HIOMT

H / 5-HT ""

5-MeOT

N-MeT

HO~N(CH')2

<0( N ~ H,CO
I NMT
NHCH,

~
H

bufotenine N
H

/ 5-MeO-N-MeT

DMT I
H]CO~N(CH]h
HIOMT /
NMT

~~
N
H

5-MeODMT

Fig. 4. Putative pathways for the endogenous production of

tryptamines from L-tryptophan. Some tryptamines can
subsequently be cyclized to form THBCs (Fig. 5).
 24

Q)~ +
N
H

biogenic carbonyl
R adduct RI substrate R RI lHBC

H3 CO 5-MeOT COOH glyoxylic acid* H3 CO H pinoline

HO 5-HT HO H 6-HO-THBC
H T H H THBC

H3 CO 5-MeOT CH3 acetaldehyde H3 CO CH3

I-Me-pinoline
HO 5-HT HO CH3
I-Me-6-HO-THBC
H T H CH3
I-Me-THBC

*decarboxylation with a-keto acids

Fig. 5. The Pictet-Spengler reaction illustrated for the production of

THBCs from biogenic tryptamines.

acetaldehyde is produced and accumulated. Disulfiram prevents

further metabolism of acetaldehyde, and MAO inhibition by
iproniazid prevents the metabolism of 5-methoxytryptamine. By
blocking these two metabolic pathways, increased levels of
acetaldehyde were reasoned to react with 5-methoxytryptamine in
a Pictet-Spengler fashion to form 6-MeO-l-Me-THBC (Fig. 6).
In that same year, Farrell and McIsaac reported 6-MeO-l-Me
THBC to be a natural component of the pineal, a cerebral gland
contaInIng high levels of tryptamine derivatives (Farrell and
McIsaac 1961). These results were never verified, and significant
analytical pitfalls have historically prevented reliable identification
of endogenous THBCs (Bosin and Holmstedt 1982). However, the
impending possibility encouraged an era of research devoted to the
 25

Iproniazid

MAO
I
5 -me thoxytryptam i ne 5-methoxyindole acetic acid

.ad re no g I om e ru 10 tropi n'

(6-MeO-I-Me- THBe)

alcohol

dehydrogenase
ethanol acetaldehyde carbon
dioxide

1
Disulfiram

Fig. 6. Mclsaac's application of the Pictet-Spengler reaction in vivo.

study of chemically based psychoses, pivoting on a dysfunctional

serotonergic system.
A major shortcoming of such thinking was in neglecting to
consider a useful function for the identified 'schizotoxins', since they
have been also found in laboratory animals and non-psychotic
volunteers as well (Riceberg and van Vunakis 1978, Raisanen and
Karkkainen 1979, Sitram et al. 1983, Raisanen 1984, Raisanen et al.
1984, Karkkliinen 1988). One hypothesis of the present thesis
suggests normal dreaming to be an appropriate and periodic
 26

expression of 'aberrant' mental activity, while an offset mechanism

may result in 'dream seepage' into the waking reality of those
individuals called 'psychotic'.
The highest concentrations of 5-HT are found in the pineal gland.
The pineal is an unpaired organ located at the center of the human
brain, where melatonin (N -acetyl-5-methoxytryptamine), a
metabolite of 5-HT, is also produced. Despite its central location in
the brain, the pineal actually resides outside of the blood-brain
barrier, and is indirectly governed by light stimulating the retina.
The retina of the eye is similar to the pineal in that both
periodically form melatonin and other methoxyindoles from 5-HT,
which also serves as a retinal neurotransmitter.
Through the retinae, light drives the periodic regulations of
biological processes into discrete circadian cycles, allowing for the
daily ebb and flow of life. Desynchronization of these cycles, such as
sleep/wake patterns, may eventually manifest as chronic and
problematic mental conditions. Since light-driven pineal activity
regulates the flux of indoleamine concentrations, various brain
activities and subsequent states of mind may be considered as
composites of these influences .
Melatonin delivers the message of 'darkness' in the scotophase of
the circadian cycle in many forms of animal life, from vertebrates
to unicellular organisms. Besides the evolutionary and bioactive
significance of melatonin and related indoleamines, melatonin has
been identified as the most potent physiological scavenger of
hydroxyl radicals known, which suggests a protective role from
radical-induced carcinogenesis and neurodegredation (Hardeland et
al. 1993).
Pinoline (6-methoxy-1,2,3,4-tetrahydro-p-carboline, 6-MeO- THBe,
5-MeO-tryptoline), a putative metabolite of both melatonin and 5
HT, has its highest concentrations in the pineal gland (Kari et al.
1983). In chickens it has been reported to fluctuate in phase with
melatonin; i.e. high concentrations during the sleep phase and low
while awake (Kari 1981). Pinoline is an exceptional neuroactive
indoleamine in that it can potentiate the activity of 5-HT in two
 27

distinctive ways: by inhibiting its presynaptic reuptake and by

inhibiting the metabolism of 5-HT by blocking the enzyme MAO-A.
The inhibition of either 5-HT reuptake or MAO-A activity are
commonly used separately as treatment modalities for depression
(Marcusson and Ross 1992, Costa and Sandler 1972, respectively),
yet no endogenous compound has been described as a unique and
definitive link in depressive disorders.
 28

2 AIMS OF THE STUDY

The primary focus of this investigation was to examine the

formations and functions of selected indole derivatives as putative
psychoactive agents.

Specific aims of this study were to:

- ascertain the potential production of THBCs from biogenic

tryptamines at physiologic pH using lH-NMR.

devise a synthetic procedure to reliably produce tritium PH]

labelled THBCs for receptor binding studies

- examine the functions of endogenous THBCs through binding

studies of the 5-HT transporter complex with [3H]pinoline.

- explore the effect of long-term exposure to psychoactive indole

alkaloids at the 5-HT uptake site on human platelets.

- evaluate the feasibility of a hypothesis linking normal dreaming

and psychosis to endogenous tryptamine derivatives .
 29

3 MA TERIALS AND METHODS

3.1 Chemicals (11, Ill, IV, VI)

The sources of commonly used chemicals are given in 11, Ill, IV,
and VI. Unless stated otherwise, chemicals were of analytical grade
or better.

3.2 Synthesis of the ~-carbolines

3.2.1 Synthesis of THBCs (IV)

The production of 1,2,3 ,4-tetrahydro-~ -carbolines (THB Cs) for

this study followed the Pictet-Spengler reaction (Fig. 5). Briefly, 1
H-THBCs and I-Me-THBCs were cyc1ized from the appropriate
tryptamine salts (tryptamine and 5-MeOT hydrochloride salts
(Sigma), 5-HT creatinine sulfate (Merck) and sodium glyoxylate
(Fluka) or freshly distilled acetaldehyde (Merck), respectively, in
an aqueous solution at neutral pH which was stirred at room
temperature for about 30 minutes (IV). An additional step of
acidification with 37% hydrochloric acid was required to
decarboxylate the 1-H -THBC-I-carboxylic acids to the desired 1-H
THBCs. The resulting reaction solutions were made basic by
dropwise addition of saturated NaHC03 (aq), and the final product
was extracted from this solution by diethyl ether. After removal of
the solvent by evaporation, the resulting THBCs were crystalized as
free bases from ethyl acetate and hexane. Yields were quantitative,
and overall recovery was typically >95%.

3.2.2 Synthesis of DHBCs (11)

Hydrochloride salts of 3,4-dihydro-~-carbolines (DHBCs) were

produced by the Mc1aren synthesis (Mc1aren 1989), and as
described for the production of 6-MeO-DHBC hydrochloride (11).
Briefly, the DHBCs . were Pictet-Spengler condensation products of
 30

diethylethoxymethylene malonate (Sigma) and the appropriate

tryptamine under very acidic conditions; i.e. using trifluoroacetic
acid as a reaction solvent. After stirring for one hour at 0 oC, the
reaction mixture was partitioned between equal portions of 1.0 M
HCl and diethylether, followed by basification of the aqueous layer
with a saturated solution of Na2C03 and extraction with CH2CI2. The
organic phase was dried over anhydrous MgS04 and filtered,
providing the free amine. Reaction of the free amine directly with
freshly prepared HCI gas , bubbled through the solution. gave the
stable hydrochloride salt in good yields, typically ~8 0 %.

3.2.3 Synthesis of [3 H]pinoline (11)

Tritium (3H) was attached at positions I and 2 of 6-MeO-THBC

(pinoline, hydrochloride salt) by a catalytic tritiation of 6-MeO
DHBC hydrochloride at the National Tritium Labelling Facility as
described (11). Specific activity was 35.5 Ci/mmol in two out of two
reactions. This product was characterized by lH/3H-NMR.

3.3 Characterization of the products

3.3.1 Thin layer chromatography (11, IV)

All thin layer chromatograms were developed on sheets of

aluminum backed silica, Kieselgel 69 F254 (Merck), using a 1: 1:1
solution of n-butanol, methanol and glacial acetic acid as the mobile
phase. After air drying at room temperature, the plates were
visualized under ultraviolet light (254 and 366 nm), and sometimes
developed in a glass chamber containing elemental iodine.

3.3.2 Mass spectrometric analysis (11, IV)

Electron impact (El) mass spectra of all products were recorded

by a VG 70-250SE magnetic sector spectrometer (VG Analytical,
UK). The resolution was typically adjusted to 10,000. The electron
 3 1

energy was 70 e V, ionization current 500 ~A and the ion source

temperature was routinely 150 QC. Samples were introduced to the
instrument via direct probe injection, where the temperature was
normally raised from 30 QC to 500 QC over 2-3 minutes. Accurate
mass measurements of the molecular ions were carried out
automatically by the data system. Perfluorokerosene was used as a
reference standard.

3.3.3 Nuclear magnetic resonance (NMR) analyses

3.3.3.1 IH and 13C-NMR analyses (IV)

1Hand 13C-NMR spectra were recorded on a Bruker AM-400

FT/ASPECT 3000 NMR spectrometer (USA). IH and COSY-NMR
spectra were recorded using a 5 mm 1HI i3C probe, operating a
400.134 MHz for 1H spectra and 100.614 MHz for 13 C spectra. The
number of data points for the 1H-NMR was 32 kW, with zero filling
to point resolution better than 0.2 Hz. A total relaxation time of 16
s for 64 scans was used for each of the spectra, with a pulse angle
of 45 Q. Decoupled 13C-NMR spectra were measured using composite
pulse sequence with 64 kW data points, 10 s relaxation delay and
90 0 pulse angle. Typically, 20-40 mg of sample were dissolved in
0.75 ml of CD30D with tetramethylsilane (TMS; 0.1 %) as an internal
standard. The detection limit of impurities was lower than 0.5
mole-% in the 1H-NMR experiments, and no impurities were
detected in samples used for further work (IV).

3.3.3.2 IH/3H-NMR analyses (11)

For the characterization of [3H]pinoline and determination of its

specific activity, IH and 3H-NMR spectra were obtained from an
IBM Instruments AF-300 spectrometer interfaced with an Aspect
3000 computer, operating at 300.135 MHz for the 1H spectra and
320.135 MHz for the 3H-NMR spectra. 3H spectra were selectively
decoupled at 5867.97 Hz. The sample was dissolved in CD30D with
 32

TMS as the reference standard. Integrations of both the proton

coupled and decoupled tritium spectra and proton spectrum
provided the mole fractions of tritiated molecular species and a
calculation of specific activity, as described (II).

3.3.3.3 1 H-NMR reaction kinetics (IV)

The Pictet-Spengler reaction for biogenic tryptamines and various

carbonyl substrates was followed over time by I H - N M R
immediately after reactants were mixed in a buffered solution at
pH 7.4 (I V). Equimolar amounts of the reactants were added, the
tryptamines as 400 III aliquots from a 0 . 16 M buffered solution and
the carbonyl substrates as 50 III aliquots from a 1.28 M solution.
Typically 3 minutes elapsed before the first spectrum was taken.
Sodium trimethylsilyl propionoate (TSP; 0.1 %) was used as an
internal standard shimming reagent for these aqueous solutions.
The decrease in intensity of the aromatic proton at position 2 for
the tryptamines was followed and compared with an increase in
the signal intensity of the resulting aliphatic protons at position 1
of the THBCs. The nM amount of THBC formed was calculated from
the integrated spectra (IV).

3.3.4 MOPAC analysis of reaction intermediates (IV)

Structural intermediates from the Pictet-Spengler reactions were

fully optimized using the semi-empirical general molecular orbital
package MOPAC 5.0, operating at the AMI level (IV). These
computer calculations were used to determine intermediate point
charges of the connecting atoms prior to cyclization, giving insight
into the necessary conditions for successful reactions.
 33

3.4 Binding studies of the SHT uptake site

3.4.1 [3H]Citalopram (VI)

Platelets were isolated from human plasma and the 5-HT uptake
activity was determined by the displacement of [3H]citalopram
(specific activity 85.5 Ci/mmol, New England Nuclear) with
unlabelled paroxetine (Beecham Pharmaceuticals) as described
(VI). This procedure was adapted from an earlier publication with
few modifications (Plenge and Mellerup 1991).

3.4.2 [3H]Pinoline (11, Ill)

[3H]Pinoline (10 nM, specific activity 35.5 Ci/mmol, 11) was

displaced from various tissues with 0.1 or 1.0 mM each of
unlabelled THBC, pinoline, 5-HT, melatonin , pargyline (all from
Sigma), citalopram (Lundbeck) and paroxetine (Beecham) in rat
brain and adrenal glands (Ill). In addition, THBC was used to
determine ICso values for [3H]pinoline in bovine cerebral and
cerebellar cortex and pineal. These binding studies followed a
previously described procedure (Kari et. al. 1988).

3.4.3 Analysis of the binding data (Ill, IV)

The nonlinear program LIGAND 2.0 (BIOSOFT, 1985), as adapted

for the Apple Macintosh from the 1980 program written by P.J.
Munson and D. Rodbard, was used to analyze and calculate the
binding maxima (B max), inhibition constants (K i), and the
dissociation constants (K d) for all of the receptor binding data
included in this study (Ill & VI). LIGAND is one of four programs in
the KELL (KINETIC, EBDA, LIGAND, LOWERY) software package.
 34

3.5 Autoradiography with [3H]pinoline (Ill)

Prepared tissue sections from rat brain or adrenals were washed

and labelled with 10 nM of [3H]pinoline at 4 0C for 60 minutes,
rinsed and air dried as described (Ill). In addition, similar samples
were prepared from bovine pineal, cerebellar and cerebral cortex.
Slices were exposed to [3H]Hyperfilm (Amersham) for 8 weeks and
autoradiograms were developed using Ilford PQ Universal
Developer.
 35

4 RESULTS

4.1 Formation of TUBCs

4.1.1 Endogenous formation of TUBCs (I, IV, V)

THBCs are probably formed exogenously and endogenously from

biogenic tryptamines through a Pictet-Spengler condensation (I: Fig.
2; V). With acetaldehyde or glyoxylate as the substrate, this
cyclization readily proceeds at neutral or acidic pH (IV: Scheme). At
physiological pH, I-Me-THBCs are not readily formed from
pyruvate in vitro. Surprisingly, formaldehyde failed to react with
any of the tryptamines to form the expected I-H-THBCs, despite it
being common knowledge from the ~-carboline literature that such
reactions take place (see V). Even under conditions of extreme pH,
both acidic and basic, tryptamines used in this study did not cyclize
to form the expected products with formaldehyde. Instead,
polymeric N -methyloltryptamines resulted (IV: Fig. 3). This led to
speculation that the cationic intermediate of the Pictet-Spengler
reaction must possess certain structural properties and electronic
characteristics prior to cyclization (IV: Table 2), These results were
compared with similar calculations of phenethylamine, a compound
known to condense with formaldehyde at neutral pH to form
tetrahydroisoquinoline (IV: Table 3). Shown in the table below are
the 'global' differentials for these intermediates; that is, the value
obtained when the point charges for the two connecting atoms are
added together. For phenethylamine, a compound known to react
with all of the listed carbonyl substrates, the global differential was
always negative. For the tryptamines, this value was positive for
the two aldehydes, and negative for the two a-keto acids (Table I).

 36

Table 1. Global differentials of intermediates from adducts and

substrates

T* 5-HT 5-MeOT PEA

formaldehyde 0.027 0.022 0.032 -0.098
acetaldehyde 0.071 0.079 0.080 -0.024
glyoxylate -0.063 -0.052 -0.530 -0.117
pyruvate -0.017 -0.00 I -0.008 -0.077

*Abbreviations; T = tryptamine, 5-HT = serotonin, 5-MeOT =

5-methoxytryptamine, PEA = phenethylamine

Overall, reactions with glyoxylate as the carbonyl substrate began

faster and continued to react at a faster rate than reactions with
acetaldehyde, and in both cases 5-MeOT cyclized faster and to a
greater extent than either 5-HT or tryptamine to form 6-MeO
THBCs (IV: Table I, Figs. 1&2).

4.1.2 Synthesis and characterization of [3H]pinoline (11)

To further investigate the properties of pinoline and related

THBCs, [3H]pinoline was synthesized (11). In the making of
[3H]pinoline a new synthetic route was devised for the production
of THBCs. This allowed for the introduction of tritium atoms to novel
positions through a catalytic reduction of the hydrochloride salt of
6-methoxy-3,4-dihydro-~-carboline (didehydro-pinoline HCI) with

tritium gas (11: Fig. 2). Also novel in this publication was the
description of a method to measure specific activity of tritiated
species by using NMR (11: Table I, Figs. 3-5).
The initial tritiated product was stored in a cold room (4 DC) as
the hydrochloride salt in an aqueous solution. However, it was
noted that the initial product had changed after six months in
storage since earlier binding results could not be reproduced. The
product from a second synthesis was subsequently stored dry
 37

under nitrogen at low temperature (-40 oC) in several vials as the

hydrochloride salt. Solutions were prepared as needed, which gave
reproducible results for up to three weeks when these solutions 0

were stored in a cold room.

4.2 Some functions of TUBes and other indoleamines

4.2.1 Binding characteristics of [3 U]pinoline (11, Ill)

In this study, [3H]pinoline (1I) was found to be a ligand for the 5

HT uptake site through receptor binding and autoradiographic
experiments (Ill). In rat brain homogenates, the specific binding of
[3H]pinoline was displaced by the following compounds: THBe >
pinoline > 5-HT > melatonin > pargyline ~ citalopram = paroxetine,
while using 0.1-1.0 mM of the displacing compound.
Among the compounds studied, THBe was found to have the
greatest ability to displace this ligand. In bovine pineal and cerebral
cortex, the IC50 values for THBC were 74 nM and 112 nM,
respectively. [3H]Pinoline was only partially displaced by the 5-HT
uptake inhibitors citalopram and paroxetine, even at high
concentrations, and the MAO inhibitor pargyline also accounted for
only partial displacement of [3H]pinoline in rat brain.
Autoradiograms showed the highest densities of binding for
[3H]pinoline to be associated with the pineal in both rat and bovine
tissues (Ill: Fig. 1 A&B, Fig. 2 C&D). Binding distributions were
nonhomogeneous in the brains of both species studied, showing
higher binding in the grey matter of cerebral and cerebellar
cortices. In rat brain slices, significant binding was seen in the
interpeduncular nucleus, median raphe nuclei, septal, hypothalamic,
pontine and central periaqueductal grey areas (Ill: Fig. 2). High
densities of binding were also observed in adrenals of rat, both in
the cortex and medulla (Ill: Fig. 1 C&D). In addition, pargyline
seemed to displace [3H]pinoline from the adrenal medula, but not
from the adrenal cortex (Ill: Fig, 1 E).
 38

4.2.2 Putative functions of endogenous indoles (I, III V)

A hypothesis has been proposed, suggesting a functional role for

THBCs (I: Fig. 2) and other tryptamine derivatives (I: Fig. 1) in the
maintenance of mental health, primarily through the regular and
periodic induction of dream phenomena during sleep (I: Fig. 3). This
hypothesis also suggests that dysfunctional dream mechanisms
could manifest in the waking phase as psychoses (I, V). The results
discussed in 4.1.1 support the suggestion that the formation of such
agents is a realistic possibility (IV).

4.2.3 Some effects of exogenous indoles (V, VI)

The oral consumption of MAO-inhibiting BCs with DMT, as In the

Amazonian beverage ayahuasca, can result in profound psychedelic
effects while the consumption of BCs alone do not (V, VI). In
addition, long term and regular use of ayahuasca was determined to
be associated with an increased binding density (B max) of 5-HT
uptake sites on human platelets (VI). B max (mean + SD; fmol/mg
protein) for the [3 H]citalopram binding site on platelets of
ayahuasca drinkers (N=13) was 993 + 238, and 724 + 164 for the
controls (N =10). This difference is statistically significant, with p =
0.006 (VI: Table 1). The mean age between these two groups was
not significantly different, nor were their values for K d.
 39

5 DISCUSSION

5.1 Formations of TUBCs (I, 11, IV, V)

THBCs are readily formed from primary tryptamines and

appropriate carbonyl substrates through a chemical cyclization
known as the Pictet-Spengler reaction (Pictet and Spengler 1911).
This reaction can occur with biogenic tryptamines at pH 7.4 (I, IV, V
and references therein) and is a convenient way to prepare THBCs
in vitro (11, V and references therein). Although in vivo conditions
may differ considerably from 'test tube' reactions, these results
(Ill) are discussed within a broader context and compared with
similarly designed in vitro experiments.

5.1.1 In vitro formation from aldehydes (I, IV, V)

While abundant evidence is available to support the fact that

THBCs can be formed from aldehydes, particularly acetaldehyde (IV
and references therein), there is little to suggest that they may be
formed from formaldehyde. Successful reactions with formaldehyde
were reported to have occurred at acidic pH with tryptophan and 5
hydroxy tryptophan (Brossi et al. 1973), though it seems unlikely
that this reaction readily proceeds with tryptamines in vivo, or
even in fermented foods. Despite this lack of evidence, the 'common
knowledge' from the scientific literature of ~-carbolines suggests
that I-H-THBCs are/can be formed from a combination of biogenic
tryptamines and formaldehyde (see V). For example, an otherwise
excellent article erroneously reported the synthesis of pinoline (6
MeO-THBC) from 5-methoxytryptamine and formaldehyde in their
Materials and Methods section (Matsubara et al. 1992), and cited a
previously published procedure for this route (Ho et al. 1968).
However, in the cited reference, Ho had actually synthesized
pinoline by the condensation of 5-methoxytryptamine with
glyoxylic acid, followed by acidic decarboxylation of the
intermediate. Pinoline and other I-H-THBCs are probably formed
 40

endogenously in this same way (Rommelspacher and Susilo 1985,

Gynther et al. 1986, IV, V).
Virtually every researcher in this field (including this author; I)
relied upon a review article of the Pictet-Spengler reaction (Whaley
and Govindachari 1951), or upon someone else's interpretation of it,
and applied this information to their own benefit without further
examination of the reactions in question. While the aforementioned
article discusses the possibility of such a formation between
tryptamines and formaldehyde, the evidence provided so far does
not support this assumption.

5.1.1.1 Formation of l-U-TUBCs from formaldehyde (IV)

In order for this reaction to proceed with primary tryptamines,

some molecular properties must be considered. With aldehydes as
substrates, a critical charge differential must exist between the
partially positive intermediate carbonium ion of the enamine and
the partially negative C-2 on the indole ring at the site of cyclization
(IV: Scheme). Relative to the ortho positions on phenethylamines,
the aromatic C-2 site on tryptamines are not as negative due to the
neigh boring nitrogen atom at position 1. To overcome this inherent
property, the carbonyl substrate must display an adequate positive
charge as a cation to form the necessary Schiff's base prior to
cyclization . In the case of formaldehyde, the carbonyl carbon
attaches to the aliphatic nitrogen of the tryptamine and then seeks
the most negative atom available, which happens to be another
carbonyl of formaldehyde for these reactions. The process is then
repeated, and a polymer is formed (IV: Fig. 3). Unlike acetaldehyde,
formaldehyde lacks a-hydrogens on a carbon atom adjacent to the
carbonyl carbon, and an intermediate cannot be stabilized through
hyperconjugation, as with acetaldehyde (Sykes 1975). For
phenethylamines, however, either of the aromatic carbons adjacent
to the ethylamine side chain are apparently negative enough to
induce cyclization to form tetrahydroisoquinolines (IV, Table 3).
 41

5.1.1.2 Formation of l-Me-TUBCs from acetaldehyde

(IV)

In the case of acetaldehyde, however, the carbonium ion

intermediates exhibit a positive charge twice that calculated for
analogous tryptamine-formaldehyde intermediates (IV, Table 2).
Also, acetaldehyde can be stabilized by hyperconjugation, making
the carbonyl oxygen a better leaving group. With phenethylamine,
the cationic charge was again double that of formaldehyde with
acetaldehyde as the substrate. In addition, the phenethylamine
intermediates demonstrated an overall negative differential of
charge (global differential, IV: Table 3) between the connecting
atoms for the aldehyde intermediates, while the analogous
tryptamine-aldehyde intermediates did not (Table 1).
These results are in line with previous findings, and lend further
evidence to indicate that primary tryptamines can readily cyclize
with acetaldehyde to form l-Me-THBCs in mammals (Beck et al.
1986 and references therein).

5.1.2 In vitro formation from ex -keto acids (IV)

The ex -keto acids, glyoxylic and pyruvic, present a special case

which illustrates another consideration; the global differential
between the two point charges of the connecting atoms (IV). This
value was positive for all tryptamine-aldehyde intermediates and
negative for intermediates of tryptamines combined with ex - keto
acids (4.1.1: Table).

5.1.2.1 Formation of l-U-TUBCs from glyoxylic acid (IV)

Tryptamine-glyoxylate intermediates gave a low value for the

cationic charge, and in all cases these values were lower than the
cationic charge for the tryptamine-formaldehyde intermediates, the
global differentials were always strongly negative (5.1.2: Table).
And in the case of phenethylamine, global differentials between the
 42

two point charges were negative, even for the phenethylamine

aldehyde intermediates (IV: Table 3). Since phenethylamines tend
to be more reactive in this regard, a negative global differential is
suggested to be a relevant factor for successful cyclization (IV). For
reactions with tryptamines, this may explain why glyoxylic acid
serves as the better substrate than acetaldehyde under conditions
of equimolar concentrations to form I-H-THBC-I-carboxylic acids,
which are readily decarboxylated to I-H-THBCs.

5.1.2.2 Formation of I-Me- THBCs from pyruvic acid (IV)

Pyruvic acid did not react well under these conditions, which is
consistent with previous reports (see Whaley and Govindachari
1951, Susilo and Rommelspacher 1988), and the global differential
was not as negative as those calculated for analogous tryptamine
glyoxylate intermediates (5.1.2: Table). Also, the cationic charges
were even less positive for tryptamine-pyruvate intermediates
than for analogous tryptamine-formaldehyde intermediates (IV:
Table 2). However, conditions of low pH can drive the formation of
1-Me-THBCs from tryptamines and pyruvate to some extent (Peura
and Nousiainen 1981, Rommelspacher et al. 1991), as acidic
conditions will tend to increase the cationic charge. Unlike the other
carbonyl substrates studied, pyruvate lacks a proton on a carbon
atom which is ~ to the carbonyl carbon. Thus, an enol must be
formed before addition of the amine to the former carbonyl carbon
of the subsequent double bond (IV). Enol formation from pyruvic
acid would also be favored by acidic conditions.

5.1.3 Biological formations (I, IV, V)

It is not only possible but probable that THBCs are formed in vivo
(Farrell and McIsaac 1961, Airaksinen and Kari 1980a, Beck et al.
1986, I, IV, V). In fact, their very ease of formation has been a
major obstacle to their confident detection and quantitation in
biological samples. The ability to accurately detect and measure
 43

these tryptamine derivatives was at one time so doubtful that the

matter remained unresolved at the end of a major international
workshop on ~-carbolines (Bloom et al. 1981).
Recent work has focused more on the presence of ~-carbolines in
foodstuffs, where the detected levels tend to be several orders of
magnitude larger than those previously reported from biological
samples (Adachi et al. 1991 a&b). Therefore, it is likely that such
compounds do exist at significant concentrations in the fermented
and processed foods we eat, and acquired through the diet. Still, the
question remains as to whether or not these analyses of food give
an accurate picture of a sample's contents before preparation and
analysis. In the absence of deuterated tryptamine standards, in
most cases, it is difficult to say with certainty if the ~-carbolines
detected are present before or after sample preparation. Even in
the absence of deuterated standards, however, well designed
feeding experiments in rats have conclusively demonstrated that
dietary manipulations can significantly affect the levels of
detectable ~ -carbo lines under certain circumstances (Adachi et al.
1993).

5.1.4 Synthesis and characterization of [3 H] pin 0 Ii n e

5.1.4.1 Synthesis of [3 H]pinoline (11)

The formation of 1,1-[3H]pinoline was accomplished by the

development of a novel synthetic route specifically designed for
this purpose (II: Fig. 2). The procedure was a modification and
extension of a published synthesis (Maclaren 1987) for the
production of stable trifluoroacetate salts of I-H-dihydro-~
carbolines (l-H-DHBCs), followed by a catalytic reduction on
palladium/carbon with tritium gas (3H2).
Mention of a [3H]pinoline product had been reported earlier
(Segonzac et al. 1987), where it was used as a potential ligand for
the 5-HT uptake site and found not to be actively accumulate in
rabbit platelets. This product was prepared as a custom synthesis
 44

by New England Nuclear, labelled with 2 atoms of tntium at C-3 on

the ~-carboline ring, and having a specific activity of 44 Ci/mmol.
Other details of the synthesis, and product characterization were
not reported. The authors noted problems with product stability
and reported initial impuntIes up to 78% when assayed by
chromatographic methods.
Attempts were made by the authors to further purify the product
prior to use, though scant evidence was obtained to verify its
identity as the final mass was reported to have amounted to only
10.6 11 g. Specific activity of the purified product used for their
binding studies was reported to have been 27.5 Ci/mmol. Despite
these problems and uncertainties, these researchers proceeded with
the experiments and eventually reported nonspecific binding for
their product. These observations are consistent with our
experiences concerning the stability of this product, as previously
described (4.1.2).

5.1.4.2 Characterization of [3 H]pinoline (11)

Measurements of specific activity are generally made by liquid

scintillation counting of a known mass of compound, mass
spectrometry, or by consideration of the IH- and 3H-NMR spectra
(Evans et al. 1985). With the availability of advanced NMR
instrumentation, it has become possible to sufficiently characterize
tritiated products by tuning the magnet 'upfield' to the resonance
frequency of tritium (11). Not only does 1H/3 H-NMR allow for the
determination of proton and tritium positions on a molecule, but it
also allows for a measurement of isotopic purity. In addition,
IH/3H-NMR may be used to determine the specific activity of a
tritiated species. This method has now been described in detail (11),
and these results are In agreement with fully characterized
products from reactions between H2 and 6-MeO-DHBC.
 45

5.2 Some functions of TUBes

5.2.1 Binding to the 5-UT uptake site (Ill, Vr

The functions of THBCs, as with many other neuroactive agents,
arise from their ability to bind to recognition sites within the CNS.
While THBCs can inhibit the enzyme MAO, particularly MAO-A, they
are not as effective in this action as their dihydro- and aromatic
homologs (Airaksinen and Kari 1980b, Airaksinen et aL. 1991).
Their primary function may be in the inhibition of monoamine
uptake (Tuomisto 1973, Komulainen et al. 1980), and particularly
the uptake of 5 -HT (Airaksinen and Kari 1991, Ill), although action
on other recognition sites have not been excluded.
Molecular modeling studies have shown that the chemical
structures of pinoline (6-MeO-THBC) and other I-H-THBCs can be
reasonably superimposed onto structures of the 5-HT uptake
inhibitors citalopram and imipramine in two different ways, and
that analogous 1-Me-THBCs do not fit well to this model. These
theoretical calculations were supported by receptor binding studies
in human platelets with [3H]citalopram, which showed pinoline to
have an ICsO of about 50 nM (Airaksinen et aL. 1991). Another
study of 13 BCs found pinoline to be the best inhibitor of
[3 H]imipramine binding in pig retina, with a K i of 170 nM
(Airaksinen and Kari 1991). An earlier study had reported an ICSO
of 320 nM for pinoline's inhibition of [3H]5-HT uptake in human
platelets (Airaksinen et aL. 1980). While unlabelled pinoline is
capable of displacing the specific 5-HT uptake inhibitors
[3 H]citalopram and [3 H]paroxetine from their binding sites at nM
concentrations, unlabeled analogs of the same were not found to be
as effective a displacers for [3 H]pinoline (Ill). All of these studies
support the hypothesis that pinoline may be a natural ligand for the
5-HT transporter, and in this way capable of modulating the
serotonergic system (Airaksinen and Kari 1991, Ill, V).
 46

5.2.2 Psychoactivity of exogenous TUBCs and other BCs

(V, VI)

As inhibitors of both MAO-A and the uptake of biogenic amines,

the BCs are able to increase neuronal activity through increased
levels of neurotransmitters such as 5-HT (Airaksinen and Kari
1980b), though such actions are not known to induce psychedelic
(also known as 'hallucinogenic') activity. The notion that some BCs
have such mind-altering effects has been propagated throughout
the literature without serious examination.
One study reported short lasting 'visual hallucinations' in mental
patients after the intravenous injection of harmine (Pennes and
Hoch 1957), with the questionable implication that some harmala
alkaloids may be 'hallucinogenic'. A later human study of the
harmala alkaloids found in ayahuasca, which also included mention
of a few other BCs, suffered from an apparent misunderstanding of
the word 'hallucinogenic' (Naranjo 1967). Naranjo's mention of the
'hall ucinogenic' properties of 6-methoxyharmalan (6-MeO-l- Me
DHBC) and 6-methoxytetrahydroharman (6-MeO-l-Me-THBC) was
most unfortunate, as he implicated such compounds as substrates
for psychoses in 'the first demonstration of an endogenous
hallucinogen' (Naranjo 1967).
The fact of the matter is that the harmala alkaloids are generally
not the 'main attraction' of psychoactive effects in ayahuasca,
though they do play a crucial role as MAO inhibitors that allow the
oral activity of DMT. Aside from tremors, nystagmus, and
sometimes simultaneous vomiting and diarrhea, harmaline alone is
essentially devoid of interesting psychoactivity when consumed
orally, even at doses considerably higher than those typically found
in ayahuasca (Callaway 1993, V, VI). Recently, this finding has been
verified and expanded upon, with the resulting synthetic mixtures
appropriately dubbed pharmahuasca (Ott 1994).
 47

5.2.3 Putative functions of endogenous TUBCs (I, V)

On the assumption that THBCs do form (or at least exist)

endogenously, it follows that they may have influence within the
CNS and perhaps even important function(s). Initially, the challenge
to measure endogenous THBCs, and other tryptamine deri vati ves,
was to identify a chemical factor for psychoses according to the
transmethylation hypothesis (Osmund and Smythies 1952, V). This
hypothesis focused primarily on the origins of hallucinatory
psychosis and sought to identify endogenous co-factors for the
etiologic basis of this disease. Based on their own personal
experiences with psychedelic substances, these and other
researchers refined the hypothesis and proposed methylated
neurotransmitters as etiological targets. This hypothesis was still
under serious investigation after 36 years (Klirkkliinen et al. 1988),
though most work in the area had ceased by the early 1980's. One
disturbing problem, aside from the technical details of detection,
was the presence of these alleged 'schizotoxins' in the biological
fluids and brain tissues of otherwise normal volunteers and
laboratory animals, respectively (see V). Apparently it had not
occurred to these researchers to suggest a normal, let alone useful,
function for these endogenous 'psychotogens'.
It has been suggested that mental states of hallucinatory
psychosis, psychedelic drugs and dreams all share common features
(Jacobs and Trulson 1979, Fischman 1983, Maurizi 1984, I, V). In a
novel approach to an old problem, psychoactive tryptamine
derivatives in the eNS of mammals are proposed herein to play a
role in the manifestations of visual and emotive phenomena during
normal dream sleep (I). Such a hypothesis would account for the
existence of endogenous psychedelic tryptamines In otherwise
normal humans and laboratory animals. The most convincing
evidence of this hypothesis exists in the psychedelic activity of the
endogenous compounds in question; DMT and 5-MeO-DMT (V).
The endogenous activity of these rapidly metabolized methylated
tryptamines is suggested to be promoted in much the same way as
 48

they are in pharmahuasca; I.e. through the regular and periodic

inhibition of MAO-A by endogenous p-carbolines (1). Appropriately,
the endogenous version of this chemical combination has been
recently termed endohuasca (Ott 1994). The hypothesis is extended
to include psychoses by suggesting that hallucinatory psychotic
episodes may result from a desynchronized dream mechanism,
where the individual essentially 'dreams' while awake .
This concept is not new, as Sigmund Freud wrote, 'A dream, then,
is psychosis' (Freud 1940). Though perhaps this was just a
paraphrase of an idea held by an early pioneer of modern
psychopharmacology, J.J. Moreau de Tours (1804-1884), who wrote,
'Insanity is the dream of the man who is awake', or maybe from
Aristotle who hinted of waking dreams as psychosis by writing, 'the
reason that we, even awake, deceive ourselves in certain illness is
the same which produces in us, in our sleep, an impression of a
dream' ( in Holmstedt 1967). What is new in this hypothesis (I) is
the identification of specific compounds already existing within the
normal CNS which have the appropriate psychoactive properties in
contexts ranging from exogenous psychoactivity, to normal mental
functions and dysfunctions.

5.2.4 Putative dysfunctions of endogenous TUBCs (Ill,

IV, V)

The aberrant formation of I-Me-THBCs as by-products of ethanol

metabolism represents a potential vector for adversely affecting
normal neuronal functions (Myers and Melchior 1977 , IV, V), since
their pharmacological profiles differ considerably from those of 1
H-THBCs (Rommelspacher et al. 1991). I-H-THBCs may act as
endogenous ligands to the 5-HT transporter and inhibit the uptake
of 5-HT (Ill), eventually resulting in more free 5-HT to interact
with serotonergic receptor sites (Briley and Moret 1993).
Conversely, a conspicuous lack of an endogenous uptake inhibitor
may result in less synaptic 5-HT through its increased uptake into
presynaptic storage vesicles, presumably for reuse or metabolism.
 49

In addition to the putative implications of molecular

pharmacology already mentioned, a recent article reported a
correlation between 'abnormal behavior' in human males with a
point mutation in the structural gene for MAO-A (Brunner et al.
1993). A complete and selective deficiency of MAO-A enzymatic
activity was found in each of the affected patients studied. While
allowing for the continued production of psychoactive indoleamines,
this form of endohuasca would not require MAO-A inhibition.

5.3 Exogenous indoles and the 5-HT uptake site (VI)

One of the salient features of humanity has been our propensity

to use drugs to modify our state of mind. From the morning cup of
coffee to the evening 'night cap', we self-medicate ourselves daily
and give little thought to the fact that our species is essentially
dependent on a wide variety of psychoactive substances. Rare
indeed is the group or individual who does not use some drug on a
daily basis.
As mentioned in the introduction, ayahuacsa is a beverage
consisting of psychoactive indoles that is consumed for religious
purposes. Since this is not an addictive substance, a characteristic
common to all psychedelic drugs, the reason for its continued use is
not to avoid withdrawal effects. In fact, ayahuasca is seldom
consumed more than once every other week under normal
circumstances. While the constituent p-carbolines are essential to
this mixture, the psychedelic effects probably manifest primarily
through the serotonergic effects of DMT on the CNS and through
increased levels of unmetabolized biogenic amines. This often
results in a profound revelation of insight into the mind/soul, the
primary effect of psychedelic drugs (V). It is no wonder that drugs
of this exceptional category have taken on religious significance.
Yet, it is surprising how little is known of the psychological and
pharmacological effects of the psychedelic drugs in humans.
Part of a recent investigation of ayahuasca was devoted to the
question of long term effects in the CNS, since thousands of people
 50

presently use this beverage and information obtained from such a

study might have applications in behavioral pharmacology,
particularly concerning serotonergic function. So far the results
have been surprising (VI). The 5-HT transporter was an initial site
of investigation due to the occurrence of this receptor on blood
platelets. A statistically significant (p=0.006) difference was found
between the control group and long time (> I 0 years) users of
ayahuasca; with a higher binding density (Bmax) of 5-HT uptake
sites in these 'tea' drinkers. This is surprising, since no other
pharmacological agent is known to significantly alter values of
B max, though the density of 5-HT uptake sites may vary
considerably from one individual to another. If true, it is likely that
other parameters of the serotonergic system are analogously
affec ted.
This brings up two interesting possibilities; either long term
consumption of ayahuasca is capable of inducing an upregulation In
the B max of platelet 5-HT uptake sites, or individuals with a
naturally high B max tend to drink ayahuasca for long periods of
time. Either possibility could have substantial impacts on the ways
we view potential treatment modalities of depression and other
affective disorders.
In the first case, an upregulation of the serotonergic system is
exactly what current antidepressant medications attempt to do; i.e.
increasing synaptic 5-HT by preventing its (re)uptake. If an
increased B max for 5-HT uptake sites were an indication of a
broader and more general upregulation of serotonergic function,
then ayahuasca could be a definitive start in understanding this
remarkable change. Since the psychological profiles of the
experimental subjects were suggestive of well adjusted individuals,
in contrast to the controls, it seems unlikely that only the density of
5-HT uptake sites has been effected. An increased B max alone
should result in less synaptic 5-HT and thus precipitate depressive
symptoms.
On the other hand, the possibility remains that long term drinkers
of ayahuasca may find relief through the tea for inherently high
 5 I

densities of 5-HT uptake sites, and that this condition allows them
to better tolerate the serotonergic effects of this mixture. This
possibility seems worthy of consideration since many" of our
experimental subjects had misused alcohol and other drugs before
they began a spiritual relationship with ayahuasca, and many more
described other symptoms of depression in their lives before
switching to ayahuasca. In this regard, we may consider that these
individuals changed their modality of self-medication for their
psychological problems, and with aya h uasca gained a distinct
advantage over their previous drug usage by the acquisition of
insight into their problems through the psychedelic effects of
ayahuasca. However, it would be unfair to ascribe all benefits to the
tea, since an individual is also taken into a new social circle which is
conducive to rehabilitation. Clearly more studies are need to decide
which of these possibilities is correct, and whether ayahuasca may
be useful in the treatment of affective disorders.
 52

6 CONCLUSIONS

Contrary to a widespread belief, biogenic tryptamines do not

readily react with formaldehyde at physiologic pH to give I-H
tetrahydro-~-carbolines (l-H-THBCs). However, due to their facile

formation from glyoxylic acid and acetaldehyde, THBCs are likely to

form endogenously from biogenic tryptamines . They may be
obtained directly from the diet as well. Molecular modeling
calculations have indicated the parameters necessary for the
successful Pictet-Spengler cyclization of biogenic tryptamines to
THBCs.

[3H]Pinoline has high affinity for the 5-HT uptake site. This and
other binding studies support the suggestion that pi noli ne may be
an endogenous ligand for this site, and indicate a modulatory role
for pinoline in the CNS and endocrine organs.

[3H]Pinoline was synthesized by a new route to THBCs, and

characterized by I H/3H-NMR. A new method to measure the specific
activity of a tritiated species by NMR is also described.

Except through their association with the visionary effects of

ayahuasca, both the harmala alkaloids and related ~-carbolines
(BCs) lack psychedelic activity. However, some BCs permit
methylated tryptamines to be orally active through the inhibition of
their oxidative metabolism by MAO-A. In the case of DMT, the
resulting psychoactivity would be psychedelic by any standards.

The psychoactivity of exogenous mixtures of tryptamines and BCs,

as in ayahuasca or pharmahuasca, is self evident. The long-term use
of ayahuasca was found to be associated with an upregulation of 5
HT uptake sites in human platelets. These findings may shed some
light on the molecular bases of both emotions and of consciousness.
 53

The confirmation of a specified role for endogenous indoleamines

(endohuasca) as agents in promoting the visions of dream sleep
and/or psychosis await further investigations. However, tne results
from these and other studies continue to support this hypothesis.
 54

7 REFERENCES

Adachi J. Mizoi Y. Naito T. Ogawa Y, Uetani Y. Ninomiya I (l991a).

Identi fication of tetrahydro-~ -carboline-3 -carboxy lic acid in foodstuffs.
human urine and human milk. J Nutr 121:646-652.

Adachi J. Mizoi Y. Naito T. Yamamoto K. Fujiwara S. Ninomiya I (199Ib) .

Determination of ~-carbolines in foodstuffs by high-performance liquid
chromatography and high-performance liquid chromatography-mass
spectrometry . J Chromatogr. 538:331-339.

Adachi J. Ueno Y. Ogawa Y. Hishida S. Yamamoto K. Ouchi H. Tatsuno Y (1993).

Acetaldehyde-ind uced formation of I-methy I-I ,2.3 .4-tetrahydro-~ - c a rbol i ne-
3-carboxylic acid in rats. Biochem Pharmacol 45(4):935-941.

Airaksinen M M. Gynther 1. Poso A. Callaway 1. Navajas C (1991). Structural

requirements for high binding affinity to the 5-HT uptake transporter for ~

carbolines. Brit J Pharmacol. December 104:370P.

Airaksinen M M and Kari E (1991). Pinoline, the natural ligand of serotonin

transporter in retina and pineal gland. Adv in the Biosci 82:239-240.

Airaksinen M M and Kari I (1980a). ~ -Carbolines, psychoactive compounds in

the mammalian body. Part I: Occurrence. origin and metabolism. Medical
Biology 59:21-34.

Airaksinen M M and Kari I (l980b). ~-Carbolines. psychoactive compounds in

the mammalian body. Part 2: Effects. Medical Biology 59: 190-211.

Airaksinen M M and W M McIsaac (1968). Indolealkylamines and behavior.

Ann Med Exp Fenn 46:367-381.
 55

Airaksinen M M. Svenska H. Tuomisto J. Komulainen H (1980). Tetrahydro-p

carbolines and corresponding tryptamines: In vitro inhibition of serotonin
and dopamine uptake by human blood platelets. Acta Pharmacal Toxicol
46:308-313.

Alander 0 R (1762). Inebriantia. an academic dissertation for Dn. Doct Carolo

Linna:o (Linnaeus) pp. I 0-11 section b ; Upsalia: (University of Uppsala).
Sweden.

Beck O. Repke D B. Faull K F (1986). 6-HydroxymeLbyltrypwline is naturally

occurring in mammalian urine: identification by combined chiral capillary
gas chromatography and high resolution mass spectrometry. Biomed Environ
Mass Spectrom 13:469-472.

Bloom F E (1982). Open questions: A summary of the workshop discussions. In :

Beta-Carbolines and Tetrahydroisoquinolines. Bloom F. Barchas 1. Sandler M.
Usdin E; eds. Proceedings of a Workshop Held at The Salk Institute. La Jolla,
California December 12 and 13. 1981..Alan R. Liss. Inc. New York. pp 401-410.

Bosin T Rand Holmstedt B (1982). Analytical pitfalls in the identification and

measurement of endogenous tetrahydro-p-carbolines. Ibid; pp 15-27 .

Briley M and Moret C (1993). Neurobiological mechanisms involved in

antidepressant therapies . Clin Neuropharmacol 16(5):387-400.

Brossi A. Focella A, Teitel S (1973) . Alkaloids in mammalian tissues. 3.

Condensation of L-tryptophan and L -5-hydroxytryptophan with
formaldehyde and acetaldehyde. J Med Chem 16(4):418-420.

Bronner H G. Nelen M, Breakefield O. Ropers H H. van Oost B A (1993) .

Abnormal behavior associated with a point mutation in the structural gene
for monoamine oxidase A. Science 262:578-580.
 56

Buckholtz N Sand Boggan W 0 (1977). Monoamine oxidase inhibition in brain

and liver produced by ~-carbolines: structure-activity relationships and
substrate specificity. Biochem Pharmacol 26: 1991-1996.

Callaway J C (1993). Tryptamines, ~-carbolines and you. MAPS Newsletter of

the Multidisciplinary Association for Psychedelic Studies 4(2):30-32.

Call away J C (1994). May we use the 'p' word now? Integration: Journal for
Mind-moving Plants and Culture 4:55-56.

Call away J C (1994). Another warning about harmala alkaloids and other MAO
inhibitors. MAPS Newsletter of the Multidisciplinary Association for
Psychedelic Studies 4(4):58.

Costa E and Sandler M, eds . (1972). Monoamine Oxidases-New Vistas. Advances

in Biochemical Psychopharmacology, Vol. 5. Raven Press, New York.

Drower E S (editor and translator) 1934. 'A Phylactery for Rue (an invocation
of the personified herb)'. Orientalia 4:324-346.

DuQuisne J T (1983). An English translation of the original Latin text, as

prepared for Prof. Mauno M. Airaksinen on 28.3.83.

Eccles J C (1992). Evolution of consciousness. Proc Natl Acad Sci 98:7320-7324.

Evans E A, Warrell D C, Elvidge J A, Jones J R, eds. (1985). Handbook of tritium

NMR spectroscopy and applications. Wiley, Chichester.

Farrell G and McIsaac W M (1961). Adrenoglomerulotropin (letter). Arch

Biochem 94:543-544.

Fischman L G (1983). Dreams, hallucinogenic drug states, and schizophrenia:

A psychological and biological comparison. Schizophr Bull 9(1):73-94.
 57

Flattery D Sand Schwartz M (1989). Haoma and Harmaline: The Botanical

Identity of the Indo-Iranian Sacred Hallucinogen 'Soma' and its Legacy in
Religion, Language, and Middle Eastern Folklore. University of ' California
publications. Near Eastern Studies; v. 21 University of California Press,
Berkeley and Los Angeles, California. ISBN: 0-520-09627-4; Library of
Congress Catalog Card Number: 82-21889

Freud S. (1940). An outline of psychoanalysis. In: "The Standard Edition of the

Complete Works of Sigmund Freud"; vol. 23. p. 172. (J Starchey and A Freud.
eds.). London: Hogarth Press.

Gunther R T (1934). 'The Greek Herbal of Dioscorides.' Oxford University

Press.

Gynther J, Lapinjoki S P, Airaksinen M M, Peura P (1986). Decarboxylation of

1.2,3,4-tetrahydro-~-carboline-I-carboxylic acids in brain homogenate and
catalysis by pyridoxal-5'-phosphate. Biochem Pharmacol 35:2671-2675.

Hardeland R, Reiter R J. Poeggeler B, D -X Tan (1993). The significance of the

metabolism of the neurohormone melatonin: antioxidative protection and
formation of bioactive substances. Neurosci Biobehav Rev 17:347-357.

Ho B T, McIsaac WM, An R. Harris R T, Walker K E (1970). Biological activities

of some 5-substituted N,N-dimethyltryptamines, a-methyltryptamines, and
gramines. Psychopharmacologia (Berl.) 16:385-394.

Ho B T, McIsaac W M, Walker K, Estevez V (1968). Inhibition of monoamine

oxidase. Influence of methyl substitution on the inhibitory activity of ~

carbolines. J Pharm Sci 57:269-274.

Holmstedt B (1982). Betacarbolines and tetrahydroisoquinolines: historical

and ethnopharmacological background. In Betacarbolines and
Tetrahydroisoquinolines, pp. 3-13, (F Bloom, J Barchas, M Sandler and E
Usdin. eds). Alan R. Liss, New York.
 58

Holmstedt BRand Lindgren J -E (1967). Chemical constituents and

pharmacology of South American snuffs. In: Ethnopharmacologic Search for
Psychoactive Drugs. DH Efron. B Holrnstedt. and NS Kline eds.. Proceedings of
a symposium held in San Francisco. Jan. 28-30. 1967 [Public Health Service
Publication No.1645] V.S. Government Printing Office. Washington. D.C .
reprinted (2nd printing) by Raven Press: New York 1979. pp. 339-373.

J acobs BLand Trulson M E (1979). Dreams. hallucinations and psychosis- the

serotonin connection. Trends Neurosci 2( 11 ):276-280.

Kari E. Tuomisto L. Airaksinen MM (1988). [3H]Imipramine binding in the

retinas of chickens and pigs. Exp Eye Res 47:679-688.

Kari I (1981). 6-Methoxy-I.2.3,4-tetrahydro-b-carboline in pineal gland of

chicken and cock. FEBS Letters vo!. 127, no. 2 pp. 277-280.

Kari I. Airaksinen MM, Gynther J, Hubtikangas A (1983) . Mass spectrometric

identification of 6-methox y- 1.2 ,3 ,4-tetrahyd ro-~ -carboline in pineal gland.
In: Recent developments in mass spectrometry in biochemistry, medicine
and environmental research 8. pp 19-24. A Frigerio, Ed. Elsevier Scientific
Publishing Company, Amsterdam.

Komulainen H, Tuomisto J, Airaksinen M M, Kari I, Peura P, Pollari L (1980).

Tetrabydro-~-carbolines and corresponding tryptamines: In vivo inhibition
of serotonin, dopamine and noradrenaline uptake in rat brain synaptosomes.
Acta Pharmacol Toxieol 46:299-307.

Luna L E and Amaringo P (1991). Ayahuasca Visions: The Religious

Iconography of a Peruvian Shaman. North Atlantic Books, Berkeley, CA.

McIsaac W M (1961). Formation of I-methyl-6-methoxy-I,2,3,4-tetrahydro-2

carboline under physiological conditions. Biochem Biophys Acta, 52:607-609.
 59

McKenna 0 J and Towers G H N (1984). Biochemistry and pharmacology of

tryptamines and beta-carbolines: A minireview. J Psychoact Drugs 16(4) :347
358.

Mclaren J A (1989). A new synthesis of 3 ,4-dihydro-~ -carbo lines (3,4,

dihydro-9H-pyrido[3,4-bJindole) Aust J of Chem 40:1617.

Marcusson J 0 and Ross S B (1990). Binding of some antidepressants to the 5

hydroxytryptamine transporter in brain and platelets. Psychopharmacology
102: 145-155.

Matsubara K. Collins M. Neafsey E (1992). Mono-N-Methylation of 1,2.3,4

tetr a hy dro - ~ -carbo lines in brain cytosol : Absence of indole methylation. J
Neurochem 59(2):505-510.

Maurizi C P (1984) . A mechanism of mania and the chemistry of dreams: A

hypothesis. South Med J 77(12):1491-1493.

Myers R 0 and Melchior C L (1977). Differential actions of voluntary alcohol

intake of tetrahydroisoquinolines or a ~ -carboline infused chronically in the
ventricle of the rat. Pharmacol Biochem Behav 7:381-392.

N aranjo C (1967). Psychotropic properties of the harmala alkaloids. In:

Ethnopharmacologic Search for Psycho active Drugs. OH Efron, B Holmstedt,
and NS Kline eds. Proceedings of a symposium held in San Francisco. Jan. 28
30. 1967 [Public Health Service Publication No.1645] U.S. Government
Printing Office. Washington. D.C.. reprinted (2nd printing) by Raven Press:
New York 1979, pp. 385-391.

Osmond H and Smythies JR (1952). Schizophrenia: A new approach. J Ment Sci

98:309-315.

Ott J (1994). Ayahuasca Analogues: Pangren Entheogens. Natural Products Co.,

Kennewick, W A.
 60

Pennes HH and Hoch PH (1957). Psychotomimetics, clinical and theoretical

considerations: Harmine, WIN-2299 and nalline. Am J of Psychiatry 113:887
892.

Peura P and Nousiainen E (1981). Synthesis and spectral data of some l-alkyl
substituted 1.2,3 ,4-tetrahydro-~ - carbolines . Acta Pharm Fenn 90: 175-178.

Pictet A and Spengler T (1911). Uber die alkaloide von isochinolin-derivaten

durch einwirkung von methylal auf phenyllHhylamine. phenyl-alanin und
tyrosin. Ber Deut Chem Ges. 44:2030-2045.

Plenge P and Mellerup ET (1991). [3HjCitalopram binding to brain and platelet

membranes of human and rat. J Neurochem 56:248-252.

Rapport M M. Green A A. and Page I H (1948) . Crystalline Serotonin. Science

108:329-330.

Riceberg LJ and Van Vunakis H (1978). Determination of N.N

dimethylindolealkylamines in plasma. blood and urine extracts by
radioimmunoassay and high pressure liquid chromatography. J Pharmacol
Exp Ther 206:158-166.

Rommelspacher H. May T, Susilo R (1991). ~-Carbolines and

tetrahydroisoquinolines: detection and function in mammals. Planta Med .
57:S85-92.

Rommelspacher Hand Susilo R (1985). Tetrahydroisoquinolines and ~

carbolines: putative natural substances in plants and animals. Prog Drug Res
29:415-459.

Raisanen M (1984) . The presence of free and conjugated bufotenin in normal

human urine. Life Sci 34:2041-2045.
 61

Rliislinen M and Karkkliinen J (1979). Mass fragmentographic quantitation of

urinary N,N-dimethyltryptamine and bufotenin. J Chromatogr Biomed Appl
162:579-584.

Raisanen M, Virkkunen M, Huttunen MO, Furman Band Karkkainen J (1984).

Increased urinary excretion of bufotenin by violent offenders with paranoid
symptoms and family violence. Lancet vol. 11 no. 4804, pp.700-701.

Segonzac A, Schoemaker H, Lee C R, Langer S Z (1987). [3 H) - 5 -

Methoxytryptoline is not actively accumulated by rabbit platelets. Naunyn
Schmiedeberg's Arch Pharmacol 336: 1-4.

Shulgin A T (1976). Profiles of psychedelic drugs. I. DMT. J Psychedelic Drugs

8:167-168.

Shulgin A T (1977). Profiles of psychedelic drugs. 4. Harmaline. J Psychedelic

Drugs 9:79-80.

Sitaram BR, Blackman GL, McLeod WR and Vaughan GN (1983). The ion-pair
extraction, purification, and liquid chromatographic analysis of
indolealkylamines in human urine. Anal Biochem 128: 11-20.

Stembach H (1991). The serotonin syndrome. Am J Psychiatr 148:705-13.

Strassman R J, Quails C R, Uhlenhuth E H, Kellner R (1994). Dose-response

study of N,N-dimethyltryptamine in humans. 11. Subjective effects and
preliminary results of a new rating scale. Arch Gen Psychiatry 51 :98-108.

Susilo R, Rommelspacher H (1988). Formation of I-methyl-p-carbolines in

rats from their possible carboxylic acid precursor. Naunyn-Schmiedeberg's
Arch Pharmacol 337:566-57l.

Spruce R (1873). On some remarkable narcotics of the Amazon Valley and

Orinoco, Ocean highways. The Geographical Reviews 1(55):184-193.
 62

Sykes P (1975). In: A guide to mechanisms in organic cbemistry, fourth

edition, Longman Group Lld., London, p 25.

Tuomisto J (1973). Inhibition of tetrahydroharman compounds of 5

hydroxytryptamine and histamine uptake in rabbit blood platelets. Naunyn
Schmiedebergs Arch Pharmacol 279:361-370.

Whaley W Hand Govindachari T R (1951). The Pictet-Spengler synthesis of

teLrahydroisoquinolines and related compounds. In: Organic Reactions
Volume 6: 151-191. Whiley: New York.
 63

ORIGINAL PUBLICATIONS

I Callaway J.c.: A proposed mechanism for the visions of

dream sleep. Medical Hypotheses 26: 119-124, 1988

I I Callaway J.c., Morimoto H., Gynther J., Airaksinen M.M. and

Williams P.G.: Synthesis of [3H]pinoline, an endogenous
tetrahydro-~-carboline. J Labelled Compds Radiopharm 31:

355-364, 1991

I I I Airaksinen M.M., Callaway J.c., Nyqvist P., Rligo L., Kari E.

and Gynther J.: Binding sites for [3H]pinoline, pp. 83-86. In:
Melatonin and the Pineal Gland: From Basic Science to
Clinical Application, Touitou Y, Arendt J and Pevet P, eds.
International Congress Series 10 17, 1993. Elsevier Science
Publications: Amsterdam

I V Callaway J.c., Gynther J., Poso A., Vepslilliinen J. and

Airaksinen M.M.: The Pictet-Spengler reaction and biogenic
tryptamines: Formation of tetrahydro-~-carbolines at
physiological pH. J Heterocyclic Chem 31 (2):431-435, 1994

V Callaway J.c., Airaksinen M.M. and Gynther J.: Endogenous

~-carbolines and other indole alkaloids in mammals.

Integration 5:1-14, 1994

V I Callaway J.c., Airaksinen M.M., McKenna DJ., Brito G.S., Grob

C.S.: Platelet serotonin uptake sites increased in drinkers of
ayahuasca. Psychopharmacology (in press), 1994
 A PROPOSED MECHANISM FOR THE
VISIONS OF DREAM SLEEP.

Callaway JC

Medical Hypotheses 26: 119-124, 1988

Reprinted with permissIOn from Medical Hypotheses, copyright

(1993), Churchill Livingston; Medical Division of Longman Group
UK, Robert Stevenson House, 1-3 Baxter's Place, Leith Walk,
Edinburgh EHl 3AF, United Kingdom
 SYNTHESIS OF [3H]PINOLINE, AN

ENDOGENOUS TETRAHYDRO

P-CARBOLINE.

Callaway J C , Morimoto H, Gynther J,

Airaksinen M M and Williams P G

Journal of Labelled Compounds and

Rad iopha r mceuticals 31: 355-364, 1991

Reprinted with permIssIOn from Journal of Labelled Compounds and

Radiopharmaceuticals, copyright (1993), John Wiley & Sons Ltd.;
Baffins Lane, Chichester, Sussex PO 19 1VD, England
 BINDING SITES FOR [3H]PINOLINE

Airaksinen M.M., Callaway J.C., Nyqvist P., Rago L.,

Kari E. and Gynther J

In: Melatonin and the Pineal Gland: From Basic

Science to Clinical Application, pp. 83-86. Touitou Y,
Arendt J and Pevet P, eds. International Congress
Series 1017, 1993. Elsevier Science Publications:
Amsterdam

Reprinted from International Congress Series 1017, copyright

(1993), Elsevier Science Publications B.Y.; Sara Burgerhartstraat 25,
P.O. Box 211, 1000 AE Amsterdam, The Netherlands
THE PICTET-SPENGLER REACTION AND

BIOGENIC TRYPTAMINES: FORMATION

OF TETRAHYDRO-~-CARBOLINES AT

PHYSIOLOGICAL pH.

Callaway J C, Gynther J, Poso A, Vepsalainen J and

Airaksinen M M

Journal of Heterocyclic Chemistry

31(2):431-435, 1994

Reprinted with permissIOn from Journal of Heterocyclic Chemistry,

copyright (1994), Dr. Raymond N. Castle, Editor; Department of
Chemistry, University of South Florida, Tampa, Florida, 33620-5250
 ENDOGENOUS ~-CARBOLINES AND

OTHER INDOLE ALKALOIDS IN

MAMMALS

Call away J C, Airaksinen M M and Gynther J

integration: journal for mind moving plants and

culture 5:1-14, 1994

Reprinted with permission from integration: journal for mind

moving plants and culture, copyright (1994), herman de vries,
editor; bilwis verJag eschenau, 97478 Knetzgau, Germany
 Platelet serotonin uptake sites
increased in drinkers of ayahuasca

Call away J C, Airaksinen M M, McKenna D J,

Brito G Sand Grob C S

Psychopharmacology (in press), 1994

Reprinted from uncorrected proof with permission from

Psychopharmacology, copyright (1994), Speinger- Verlag GmbH &
Co. KG; D-14197 Berlin, Germany
I

 Medical Hypotheses .

Mnll~'; IJ II Y I~'lh ~' '''''' ~ (I ()XX) ~f'I. III}-I:-'

1.lIn"m an U ruup UK IYXH

A Proposed Mechanism for the Visions of Dream

Sleep

J. C. CALLAWAY

School of Pharmacy, University of California, San Francisco, California 94143

Abstract - The visions of dream sleep are suggested to occur through a dream
mechanism which implicates tryptamine derivatives as endogenous paychedelics. The
hallucinations that occur in some schizophrenic syndromes are also proposed to occur
through a similar, though desynchronized, mechanism. These compounds occur in the
human pineal gland and are regarded as neurotransmitters or neuroregulators . A protocol
for experimental verification is suggested.

Introduction
Much has bcen wri!!en concerning various slates
of consciousness precipitated with and without
the assistance of drugs. though most of the
scientific literature distances itself from the
psychedelic experience by suggesting it as a
model of induced psychosis (i .e . psychotomi
metic) or other undesirahle state of mind . Few
have considered this experience as a normal
occurrence (12) . However. it has heen suggestcd
th<lt the human urge to dcliberately illter
consciousness is <IS innate as <lny natural drive
(35). This idea is extended by the suggestion that
through dreams the mind experiences altern<ltive
states of consciousness on a regular basis. and
gains perspective and insight into waking reality.
Drcam phel10menil are tr<lnscultural. prcdate
literature and occur in virtually all humans (and
perhaps other animals) several times a night. Yet
dreams arc perhaps the least understood facet of
human experience.
Humans spend approximately one third of
their lives asleep. During this period of rest and
rejuvenation. less than one fourth of the time is
spent in the drcam statc commonly referred to
as rapid-eye-movement (I{EM) sleep. or REMS.
REMS is the last of five yualiWtively different
stages of sleep th<lt Ciln be chilracteri zed by elee
troencephlographic activity. Although the corre
lation hetween I{EMS and dream sleep i, strong
it is not absolutc. ( ,",or revicws of dre ,'m sleep
sec references 7 and 14). The I{EM stilge usuillly
increases in length during the slccp cycle . often
reaching a pcriod of approximiltely two hours
prior to waking. Dreams also increase in length
ilnd intensity throughout the sleep cyc le . The
most vivid and emotionally charged dreams also
tcnd to occur just prior to waking. These MC the
dreilms best remcmbered in the wilking phase of
ths hum an circadiiln cycle.
This article suggests an outline of a dreaming
mechanism and cites endogenous compounds

119
120
that possibly play a role in REMS, and more
specifically, the visual phenomenon of lucid
dreaming. Such compounds could also account
for other alternative states of consciousness, such
as experienced in certain meditative practices,
and might be psychedelic when administered
during the waking phase . They must be
produced endogenously during sleep , with their
dream-inducing qualities being dose dependent
and relatively short acting (IS minutes to two
hours). Dose dependency is indicated by the
gradual increase of REM activity throughout the
sleep phase. when most of the neurotransmitt ers
are produced to sustain longer episodes of
REMS. In as much as humans dream periodi
cally, these compounds would also be involved
in a mechanism of dream initiation/inhibition to
start and stop the dream process . In this way the
dream state does not dominate the sleep phase
and allows for a certain amount of vigilance to
be maintained. Though some overlap may be
necessary for normal "day dreaming", excessive
seepage may occur into the waking phase
without such a periodic regulatory mechanism.
Such an overlap may occur in some schizo
phrenic patients, where a desynchronized dream
mechanism allows the intrusion of dream chemi
cals into the waking state (20). Such chemicals
have been proposed in the transmethylation
hypothesis (26). Rather than limiting this mech
anism to abnormal behavior, the transmethyla
tion of specific endogenous compounds during
sleep may actually be an important part of
normal mental health within a properly func
tioning dream mechanism, and not merely the
harbinger of mental disease. beta-Carbolines and
tryptamines are two classes of compounds impli
cated in this balance of dreaming and insanity .
beta-Carbolines (harmala alkaloids) are alka
loids found in many plants and animals. They are
thought to occur in humans as condensation
products of indolealkylamines (tryptamines) with
aldeh ydes (formaldehyde), formed through enzy
matic reactions with S-methyitetrahydrofolate (S
MTHF) and/or S-adenosyl-I-methionine (SAM),
and catalyzed by N-methyltransferase (NMT) (I,
6, 10. 30, 34). This condensation is followed by
a less understood cyclization . In addition, beta
carbolines have been proposed as endogenous
ligands to the so called benzodiazepine receptor
(19,23), suggesting a link to anxiety release and
control.
Some of the beta-carbolines have demon
strated psychoactive properties in humans when
MEDICAL HYPOTHESES
taken in their pure form (32). but are qualita
tively different in their effects from LSD or
mescaline. Naranjo (24. 2S) stated that distortion
of form. depth-movement perception and color
enhancement were not observed in a series of
beta-carbolines. The most frequently occurring
phenomena included . "superimposition of images
on flat surfaces and viewing scenes simul
taneously with an undistorted perception of
surrounding objects". Abundant bright and vivid
colors were reported with eyes closed . ~ccording
to Naranjo (2S) the characteri stic effects of 0
methox y- belil-carbolines were considcrcd . "to be
of a less hallucinogenic nature being more
akin to a state of inspiration and heightened
ir1trospection
NaranJo (2S) also noted a recurrent visual
phenomena of. "rapid lateral vibration in the
vision field." Harmaline has been found to
produce a generalized tremor at a frequency of
eight-12 Hz in many animals (10). This same
band of frequencies has been designated as alpha
brain waves in humans. These waves occur
hormally when the eyes are closed in a waking
state and during REMS, but not during the other
four stages of sleep (7). The harmaline induced
vibratory phenomenon of the visual field is inter
esting in the context of REMS. where the rapid
eye movements echo a deeper process in sleep.
Anthropological data suggest that harmala alka
loids (beta-carbolines) may be more than merely
hallucinogenic in actual usage. These compounds
have traditionally been used in preliterate soci
eties to produce out-of-body experiences clair
voyance. simultaneous group visions. remote
viewing and divination (IS. 24) .
About 10 different beta-carbolines have been
found in mammalian tissue (I). Although their
metabolic precursors are naturally available. the
exact in vivo biosynthetic pathways involved
have yet to be definitively established . Water
soluble metabolites of endogenously produced
beta-carbolines have been identified in human
urine and there is increasing evidence to suggest
that some of these compounds are formed under
special physiological circumstances, such as after
alcohol consumption and perhaps at other times
(1. 2. 28. 34). beta-Carbolincs are found in
human plasma and are highly concentrated in
platelets (33). 6-Methoxytetrahydro-beta-carbo
line, also called 6-methoxytetrahydronorharman,
has been found in the human pineal gland (29)
and retina (16), possibly acting as a neuromo
dulator or neurotransmitter, and has been
A PROPOSED MECHANISM FOR TIlE VlSIONS OF DREAM SLEEP
named pinoline. Like most beta-carbolines.
pinoline is a typical serotonergic compound. a
property shared with psilocybin and other
psychedelic tryptamines. It inhibits the enzyme
monoamine oxidase-A (MAO-A) and has the
unique property of increasing concentrations of
brain serotonin (10) . Since pinoline and other
endogenous beta-Carbolines inhibit MAO-A.
they can competitivel)' inhibit that enzyme while
other psychoactive but metabolically more labile
compounds. such as tryptamines. act centrally to
promote the visions of dreams. A synergistie
relationship between ingested tryptamines and
beta-carbolines has been auggested (21. 22) .
Dimethyltryptamines have not been definitively
established to occur endogenously in humans.
thoug.h the in vitro evidence is hig.hlv suggcstive
(-I . 5. :11). This lack or confirmation is not
~urprising wh<.:n onc considers the minute
(on(<.:ntr,ltions involved. sensitivity of deteLtion
methods ,lOd instability of endogenous tryptam
ines (11) . (Sce reference 31 for a review of
endogenous paychedelics) . The time of sampling
during the circadian cycle may also be crucial in
detecting psychodynamic tryptamines (18).
During the sleep phase. melatonin is derived
from serotonin. which is in turn metabolized
from the amino acid tryptophan. 5-Methoxydi
methyltryptamine and DMT. both potent
psychedelics. have also been proposed by Barker
(5) to derive from tryptophan through SAM and
NMT (sce Fig. I) . Melatonin. like serotonin. is
produced primarily in the pineal gland. Its
~ .O:JNH,
~ Decarboxylation
TRYPTOPIWI TRYPTNIINE
I. SAIl
2. NHT I
'"
DIHETHYLTRYPTNIINE
Figure 1
121
generation seems' to be inhibited by light
entering the eye (3. 8, 18) . Generation of mela
tonin may begin soon after the eyes have closed.
as in the onset of sleep and perhaps in some
meditative states . During the sleep cycle of most
animals studied . including humans, the levels of
serotonil1 decrcased while the levels of melatonin
increased (18). According to Ebihara (8). this
decrease in serotonin is partly due to its trans
formation to melatonin (sce Fig. 2). Endogenous
beta-Carbolines are also proposed to derive from
serotonin through the increased activity of NMT
and 5-hydroxyindole-0-methyltransferase (5
HIOMT). both pineal enzymes of the sleep
phase (I. 6. 10) . Pinoline and mel,ltonin were
found to occur in neurly simililr concentrations
in the human retina. The origin of retinal pino
line seemed to he mostly pineill. with only a frac
tion being locally synthesized (17). All of the
chcmical structures shown in Figure 2 hilve been
identified in the humiln pineal. The intermediate
structure in brackets is one of two proposed by
Ho (10). The solid arrows denote pathways
which are known to occur in vivo. while the
broken arrows have only so far been demon
strated to occur in vitro .
Figure 3 represents a proposed dream cycle.
As melatonin production increases in the sleep
phase. the brain descends through the four
stages of sleep . beta-Carbolines and dimethyl
tryptamines arc derived from tryptamine. and
when enough arc produced. the mind enters
dream sleep. This is rcflected by the brain as
HO
~
~~
H
SEROTONIN
5-HETHOXY
DIHETHYLTRYPTNIINE
122
PINOLINE
Figure 2
REMS . Pinoline incrcases brain serotonin levels .
especially at low concentrations (2. 10. 33).
which eventually inhibits dream sleep by taking
the brain back through the four sleep stages.
Before the cycle reaches the waking phase. N
acetyltransferase (NAT). NMT and 5-HIOMT
work to metabol ize the excess serotonin to
melatonin. pinoline and perhaps other tryp
tamine derivatives. The cycle then repeats and
its period lengthens throughout the night. The
longest and best remembered dreams occur Just
prior to waking from the cumulative concen
trations of psychoactive tryptamine derivatives
which have become active through progressive
inhibition of MAO-A by pinoline and/or other
beta-Carbolines. Other circadian mechanisms
come into play before waking so that the dream
cycle does not overlap into waking conscious
ness. It is not unlikely that beta-Carbolines
and/ or other tryptamine derivatives act centrally
to induce the visions of dreams . Although Ho
(10) was speaking in terms of psychosis. the
following applies with fcw modifications. "It is
quite possible that a psychotic state (;()uld result
from a shunt of serotonin metabolism to a
MEDICAL HYPOTHESES
I eye 1 iz.,1t 10n
HO -J.
'OcD N
H
H
pathway that occurs and leads 10 accumulations
of abnormal amounts of such potentially psycho
tomimetic compounds ."
This dream cycle hypothesis could be tested in
a couple of e xperiments . Onc would be to
monitor the ~erum and/ or cerebral spinal nuid
levels o f beta-Carbolines throughout the circa
dian cycle . with expectations of higher concen
trations during sleep a nd the REM stages of
sleep. A 1110re ambitious procedure would be to
determine beta-Carboline levels in normal and
schizophrenic subjects before. during and after
REMS deprivation . When normal individuals arc
deprived of REMS. a rebound of REM activity
occurs in subsequent nights of undisturbed sleep .
Schizophrenics show no such rebound. Therefore
onc might expect to sce elevated beta-Carboline
levels during rebound in normals. with schizo
phrenics maintaining their normal predeprivation
levels. Morning levels of pilloline in normal and
schizophrenic patients were found to be equiva
lent after normal sleep (2<j). Animal studies
suggest that pineal levels of pinoline and mela
tonin arc of the same order and change in
concert during the circadian cycle (13). Mela
A PROPOSED MECHANISM FOR THE VISIONS OF DREAM SLEEP
S.rotonin lncrea ...
""0<0"'1 ,,,,,....
Sotrotonln d.et
Hel_to,..1n lncr
1
!'kl.tonln prod... ct 10"
lncr . . . . . "Ilk ,hip
Du , ItOP MAO inhiblt10n OCCIH,
DatA." CYCU
Serotonin co~.t
wllh p.ycho.ct I ....
ttypt lne d, r lvu I"
Wake Cych
Figure 3
tonin administered before sleep decreases the
time of sleep onset and increases the incidents
of vivid and colored dreams (27). and could
serve as a precursor to psychoactive beta
Carbolines.
It should be mentioned that this hypothesis is
not the currently accepted view of a mechanism
for REM-dream sleep. In a recent article (Y).
acetylcholine was the neurotransmitter impli
c<Jted as being responsible for initiating and
maintaining REMS. It is quite likely that acetyl
choline. along with several other neurotransmit
ters. plays a major role in REMS. yet the actual
catalyst for the dream state itself may be quite
different in nature and function than the trans
mitters involved in the physiology of REMS .
Conclusion
This article speculates on the endogenous
production of psychedelic compounds to account
123
"'AX[ CYCLE
for dream phenomena through a metabolic
dream cycle. and posits that such a mechanism
may be discrete from the physiological symptom
SL[EP onr.
atology observed in the rapid eye movements of
REMS. Wilh tryptamines and b~t<J-Carbolines
proposed as a seareh targets. it may eventually
beeome possible to find "the kind of stuff that
dreams are madc un."
Acknowledgements
chro",." pllWl.i ~f.
cub<l11n.c prod",ctlon The In vOI luahlr cOlllrinulinns from Daviu Pale. Richil((J
Scymour ilnd Akx;tnlkr Shulg,in were vit.1I 10 (he evolutiou
of thi:-- p i lPI.."L
References
J. Alraksincn M M. Kari I. bC(;J,C;lrholinc:-- . P:--ychoactive
CompoulH.h in Ihl...' Mammali.-tn Uolly. P;lrI I. OccurH.'nCI'
anll Mcl;1I11111:--m . Mc.:di(.a J BIt'lo~y SI.): 21. II.}X I .
ftH~lh
Dr occloIr
uninhibited I(tlon of
2. Airak:--incll M M. K~ri I. hl...'taCarholincs. P'y(.huiKtlv(.'
bltl c.rbolinl. C(lmrxlunlh in lhc M;lInmalian L3oLly. PiJrI 11. Efkt.:h .
,,,d/or other layctlet Mcuicd tli"h)~y )'i: 1'111. I~XI.
ut ,,,. I typt lnl
3. AkcNeUI T. hoher~ J E. ",,"nil Y. Wellerherg L.
11.}71.J . MdiJtonin Fxu(..'lion. Bnuy Tcrnj1\:raturc: anLl
/ "Suhlcctiv1..' Arou:-.al During. 64 Hours or Skc:p Dc:priviJ'
lion . Psy dlOncuroellllouill()lo~y .a : 21Y, 197Y.
4. 11Jrker SA. Monli J A. Chri"ian ST. N.NDlmelhyl
IryrLlllline : an .... ndll~eIlClu:-- Hidlu(ino!!en. International
RcvlCw III Ncurohll,I\I~y 2~: X.,. P)XI .
5. Barker SA. MOnli J A. (,h"'''Jn ST. MelJholism of
the H.. llu("H,~cn N . NDlmethyltryrlaminc in ROIl Hr'lin
HlIll\ogc:nall.::--. Bltu.: hcmKal Pharmacology 2t): I04 l ).
l'iXII.
h. Bu(kholtz N S. Minircvicw. Neurohiology of hctiJ
Carholinc::-.. Lift: SClcn(c:s 27: XYJ. IYXO.
7. ('arlwrighl R 0 (EtL) . A Primer on Sleep Jnu
Dr1..'aming. ALlLlison Wcs\c:y PuolishlOg Comp;..IOY. Mcnlo
Park. Ca"forma. IY7X .
X. Ehihar" S. M,lrks T. Huuson 0 J. Mcnaker M. Gene!ic'
Conlrol of MclJlOnin Synlhesis in Ihe Pineal Glanu of
Ihe Mouse . Science 231: 4~1. IYX/).
'I. Gillin J C. Si"Ir"m N. Janowsky O. Riseh C. Huey L.
Sloreh F I Cholinergic Mech,lnisms in HEM Sleep. In
Sleep: NCufolfilnsmilters .. nll Ncurofcgulators. (A
Wa"'luier. J M Gaill'lru. J M Monli. M Rauulovacki.
cLl:-.) Raven Press. Ne w York. IYH5.
10. Ho B T. Pharmacolog.i(,al altll Uiol:hemic:d stuuics wilh
hC:lilC.. rholine Allalog!\. Current Deve!opmt.:nts in
P:-.yc..:hoplwrl1lil(ology .t : 1)2. 1979.
11. Hryhorcwk L M. Raillcy J M. hoh"m C E. Novak
EA . A New Melaholic 1';)lhway for N .NOimclhyllryp
laminc .. lIiological I'sychialrY 21 : X4. I~X/) .
12. Jilcohs U L. DrciJm~ anLl Hallucinalions : A Common
Neurochc:mical MCl:hanism MeLlialing their Phcnome
11OIogical Simi\;.Hities. Ncuroscienc.:c & Uiohchavioral
Reviews. 2: W. 1'I7X.
13. Kari I. /)-Melhoxy-I.2.3.4-lelrahyJro-oela-earo"line in
Pineal Glanu of ('hick en anJ Cock. FEBS Lellers 127.
2: 277. I'IXI.
14. Lallcrge S. LUl'iu Dre~ming. Ballanline Books. New
York. IYX,).
15. Lamo B F (Eu.). Wizaru of Ihe Upper Amazon: The
124
Story of Manuel Co rdova Rios. Houghton Mifllin .
Boston. 1975.
16. Leino M. &-Methoxytetrahyurobeta-carboline and Mela
tonin in Humane Retina . Experimental Eye Research
37: 325. 1984.
17. Leino M. Airaksinen MM. Gynther J . 6-Methoxytetrah
ydro-beta-carboline in the Retina of Rabbits and Pigs.
Experimental Eye Research 36: 135. 1983.
HI. Lcwy J A. Effects of Light on Human Melatonin
Production and the Human Circadian System. Progress
in Neuro-psycho-pharmaeology and Biological Psychiatry
7: 551. 1983.
19. Lippke K P. Schunack W G. Wenning W. Muller W E.
betaCarbolines as Benzodiazeplne Reeeptor Ligands .
Journal of MedIcinal Chemistry 20: 44~. I~IU.
20. Mau" ,i C P. The Anatomy and Chemistry of Halluci
n<.Jlions <Jnt! a R<Jlion<.J1 Surgical ArproiJch 10 Ihe Trc;.J(
ment of 'ome SchizophrenIC Syndrome,. Mollical
Hypoth"is 17 : 227. I~H5.
21. McKenna DJ. Towers G H N. Biochemi,try and Phar
macology of Tryptamines and heta-Carholine> A Mini
review. Journal of Psychoactive Drug' 10(4) : .1~7. I~K~ .
22 . McKenna 0 J. Towers G H N. Ahholl F. Mon,)amine
Oxiu<Jse inhihilOrs In South Amcric<Jn HaUuc.:inogl.:'nlc
PI ... nl ~; Trypt<.Jmine anu hetaCarboline ConslilucnlS of
ayahuasca. Journal of Ethnopharmacology Ill: 195. l~x.I .
23. Muller W E. Fehske K J. Borbe H O. Wollert U. Nanz
C. Rommelspacher H. On the Neuropharmacology of
Harman and other beta-Carbolines. PharmaC<llogy
Biochemistry and Behavior 14: 693. 19!51.
24. Naranjo C. Hallucinogenic Plant Use and Related
Ind igenous Belief Systems in the Equado"an Amazon.
Jou rnal of Ethnopharmacology I: 121. 1~7~ .
25. Naranjo C. Psychotropic Properties of Harmala Alka
loids. p 3Wi in Ethnopharmaeo logic Search for Psychoac
tive Drug, . (D H Efron. 0 ti<)lm"edt . N S Kline. eds . ).
US Public Health Service PublIcation # 1045. Wa , h
ington. D.e. 1907.
26. Osmond H . Smythies J . SChizophrenia: A New
Approach. Journal of Mental Science 9M: 31)9. 1'.152.
MEDICAL HYPOTHESES
27. Pavel S. Goldstein R. Petrescu M. Pope M. Melatonin.
Vasotocin and REM Sleep in Prepurbal Boys . In
Advance, in the Biosciences. Vol. 29. Melatonin -
Current Status anu PerspectIve s. Proceedings of an Inter
national Svmposium on Melatonin . Breman. FRG. 19HO .
(N Birau. W Sehlool. eds.). Pergamon Press. New York .
19MI.
2M. Pcrua P. KMi I. Airaksinen M M. Identi~cation hv
SelectIon Ion Monlto"ng of I-Met hyl-I .2 ..1.4.tctrahvdru'.
heta-carbolinc in Human Platelets (.Inu Pla\mtJ ..Jflcr
Ethanol Intake . Biomedical Mass Spectrometry. 7: 55 3.
19MO .
29. Rimon R . Airakslnen MM . Kari I. Gynther J. Vena
laine!) E . Heikkila L. Ryyppo J. Palo J . Plno,,;'e. a heta
C;'Hholinc In the Scrum JfHJ C .:rl'hro"rJl n<.l1 FllilU of
Palients wllh SchlzortHcnlil. Ann,lIs of Clinical Rc~earch
10: 17 1. I'JH-I.
)0 . Hommebp<lchcr H . Su:-.llo H, Tctr;Jh~t!roi~llqUln ol ,"cs
. (l nt! hcta-Carhulincs : PUiatlvc N,lIural Sun'I'''l(c~ In
Plan\) ;Jnd MJOlm..lb , Progrc\:-. In Orug RI."''IcJrch
~~: 41). I'IK)
31. Rlh.engJrh:n If , Frlcuhoff A J. A ReView (If RCI.'I.'nl
SllII...ilc s \.r Hall\lclno~cn\ Ftlrml.:u hy Mclhyl.lllon of
Ncurolr;.an\mil(c r~ or Hd;lIcti ~uh,tiJncc\, SChlzophrcnl<l
L!ullelJn 2: '10. IY70.
.12. Shulgln AT . Pwlill:-\ of P:-. yc he<.klic..: Orug~: Hilrrn;llJnc .
Journal of P,ychedelic Drugs ~ (I): N. 1~77.
33. Taylor D L. Tan"y W. Cook J M. Ho 8. T. Ev;,ju"tlon
of Two Distinctive nel;'I' CiJrbollnes on Serotonin
Bintiing In Human Pl ah:lcls, Research CommunlCiJlions
in Clinical Pathology and PharmaCOlogy 47 : 1.1.1. 14K5.
.14. Torn:iliL'\ 1. (iucrln M. Prevlcro . A . f)t:~ SlruCluro
Simplcs ,lU X POlcrlllalilC S Philrrnacok'glC..jUC\ l:.1l'\et.'s : k~
beta-Carholines (4H-py"do 13.4 bl-indole). Origines.
Synlhc\cs. Propcrll:tes Biologiquc" 13lochimle h7: 1.)21.).
I"X) .
35 . Well A W (cd .). The Natural Mind : A Way of l.ooklng
al Orug\ anu High<.:r Con\ciou\ncss, 13U\lon: Houghlon
Mifilin. 1')72.
11

Journal of Labelled CompoUllds and Radiopharmaceuticals- Vol. XXXI. No. 5

Synthesis of [3H]Pinoline, an Endogenous Tetrahydro-l3-carboline

J.C. Call away 1,2, H. Morimot0 3 , J. Gynther 1 , M.M. Airaksinen 2 and

P.G. Williams 3

1 Department of Pharmaceutical Chemistry, 2Department of

Pharmacology and Toxicology, University of Kuopio, POB 1627,
70211 Kuopio, Finland. 3National Tritium Labeling Facility,
Lawrence Berkeiey Laboratory, Berkeley, California 94720

SUMMARY

Pinoline (6-methoxy-1,2 ,3 ,4-tetrahyro-9H-pyrido[3,4-b]indole) is an

endogenous l3-carboline thought to be synthesized in the pineal gland from
serotonin . It is a strong inhibitor of MAO-A and can displace [3H]citalopram
from the 5-HT uptake site on human platelets in nM concentrations. The
hydrochloride salt of 6-MeO-DHf3C (6-methoxy-1,2-dihydro -9H-pyrido[3,4
b]indole), also a new compound, was synthesized as the immediate precursor
to the title compound.

Key words: Tritium NMR , pinoline, l3-carboline, tryptamine, endogenous

INTRODUCTION

I3-Carbolines (I3Cs) are of interest since they occur in many plants and
animals (4) . Synthetic analogues can precipitate reactions from anxiogenic
to anxiolytic, depending on substitution, and interact with several
neurotransmitter systems (10) . They inhibit monoamine oxidase-A (MAO-A)

0362 -4803 /92/050355 -10$05.00 Recei ved 11 November. 199 1

199 2 by John Wil ey & Sons. Lld . Revi sed 14 February, 1992
356 l.C. Callaway er al.

(5) and can interact with the benzodiazepine binding site as antagonists,
agonists and inverse agonists (6). This certainly hints of some
neuroregulatory role in stress modulation for endogenous I3Cs. In animals
they are thought to form as condensation products from tryptamines and
aldehydes (8) and/or keto acids (7) . Recent research has shown that 13
carbolines do not occur as artifacts in analysis or as dietary components (1).
About half a dozen different I3Cs have been identified in humans, t~ough

little else is known of their origin or function .

Condensation of endogenous tryptamines with formaldehyde, formed
enzymatically from 5-methoxytetra'hydrofolate (5-MTHF), result in I3Cs
unsubstituted at C-1 (Fig . 1). With acetaldehyde, from the metabolism of
ethanol for example, products methylated at C-1 result. There is current
speculation as to their role in alcoholism (2 ,11).

5 4

10cCN2
6 ~

n- carbollne 3,4- dlhydro-n-carbollne HCI 1,2,3,4- tetrahydro-n-cnrbollne

nc DHnc HCI THnc

Figure 1

Tetrahydro -l3-carbolines (THI3Cs) can be produced synthetically by

catalytic hydrogenation of dihydro-l3-carbolines (DHI3Cs). Essentially no

work has been done on DHI3Cs which are unsubstituted at C-1, perhaps due to

their lack of stability and subsequent availability. The trifluoroacetate of

DHI3C has recently been reported as a fine crystalline solid , produced in good

yield (9), though these salts are not appropriate for receptor binding studies,

Therefore we have developed a method for preparing the hydrochloride salt

of dihydro-pinoline for biological investigations and as a precursor to

[3Hjpinoline (Fig, 2),

Synthesis of [JHJ Pinoline 357

HyDE' _
+
C (co, El),

5-melhDHy-lryp IlImlne

1. f ,COOH

j 2 . HCI (g)

2. Pd/C
40'4 "-T

60'7. "-H

('HJptnollne dlhydrD-ptnollne HCt

figure 2

EXPERIMENTAL
Materials
Solvents were of analytical grade or better. Oiethylethoxymethylene
malonate and 5-methoxytryptamine were purchased from Sigma. Ethanol 200
proof and 10% Pd/C were purchased from Aldrich.

Synthesis of 6-methoxy-3,4-dihydro-B-carboline hydrochloride

A 1.1 molar excess of diethylethoxymethylene malonate, 1.041 g (2.89

mM), was added to 500 mg (2.63 mM) of 5-methoxytryptamine and stirred for
one hour at 95C in a flask heated by a water bath. The reaction was cooled
to room temperature and ethylacetate was added to give a total volume of
20 mL. This solution was gravity filtered, the solvent and excess malonate
were removed in vacuo, leaving 950 mg of the crude enamine. Without
further purification, the remaining oil was dissolved in as little
trifluoroacetic acid as possible (1-2 mL to 1 g of crude enamine) while
stirring in a flask on ice for 2 hours, forming the trifluoroacetate salt of 6
MeO-OHI3C. Excess acid was partially removed in vacuo at room temperature.
358 l.C. Cal/away et al.

The crude product was partitioned between 1 M HCI and ether. The aqueous
phase was made basic by drop-wise addition of sat. aq. Na2C 03 and extracted
with CH2CI2. The organ ic phase was dried over anhyd . MgS0 4 for 1 hour. This

solution was gravity filtered and the total volume was reduced in vacuo to
100 mL. the released vacuum was filled with nitrogen gas rather than

ambient atmosphere to avoid decomposition of the free amine.

HCI gas was prepared by dropping conc. H2S04 into a flask containing
conc. HCI and a small amount of NaC!. The mixture was gently heated. and the
generated gas passed through a refluxing column and a column of CaCI2
before it was bubbled into the dry CH2CI2/ 6-MeO -DH13C solution for about
30 seconds. The product was recovered by removal of the solvent in vacuo.

and crystallized from dry ethylacetate (MgS04) while using a minimal

amount of hexane to induce precipitation.

Synthesis of [3H]pinoline hydrochloride

20 mg (0 .08 mM) of 6-MeO-DH13C hydrochloride was dissolved into 3

mL of 200 proof ethanol in a 15 mL reaction flask. giving a clear yellow
solution , and frozen in a bath of liquid nitrogen under high vacuum (10- 12

Torr) . The reaction vessel was purged with nitrogen gas while the sample
slowly thawed to room temperature. After the third evacuation, tritium
(from a heated . uranium hydride source). was introduced into the reaction
vessel until the pressure reached 730 Torr, then 20 mg of 10% Pd/C was

added to the solution from within the system. After one hour of stirring at
room temperature the reaction solution appeared lighter in col or . The
reaction was terminated after a total time of 1.5 hours. The product was

immediately converted to the hydrochloride salt by the addition of ethanolic

HCI. thus avoiding possible loss of the product via sublimation. The solution
was fi ltered to remove the catalyst; the filtrate was a pale-yellow solution.
Ethanol, excess HCI and labile tritium were removed by lyophilization of the

filtrate. A pale yellow solid remained which was dissolved in methanol and

again lyophilized .
Synrhesis of [JH] Pinoline 359

Characterization of the Products

High resolution mass spectra were recorded using a Trio-2-VG Masslab

(Manchester U.K.) mass spectrometer. The electron energy was 70 eV, ion
source temperature 200C, and ionization current 200 /lA. Samples were
introduced by a glass holder with a direct insertion probe. Accurate mass
measurements of molecular ions were carried out automatically wjth the
data system. Perfluorokerosene was used as the reference compound.
NMR spectra were obtained from an IBM Instruments AF-300
spectrometer with an Oxford magnet of field strength 7.05 Tesla (70 K
Gauss) interfaced with and Aspect 3000 computer with Digital Phase
shifting 1 0 resolution) using a 5 mm 3H/' H dual probe. All samples were
dissolved in CD30D with TMS as the reference standard; SF .. 300.135 MHz
and SR .. 4550.59 Hz for the proton spectrum; SF .. 320.135 MHz and SR=
5438.10 Hz for the tritium spectra, which was also selectively decoupled at
5867.97 Hz.

6-MeO-OHI3C hydrochloride. The yield was 512 mg (83 %), of a pale

yellow crystalline powder which decomposed at 200C. High resolution MS:
m/z 201 (12 %), 200 (74 %),199 (100 %), 184 (14 %),172 (8%), 156 (11%),
130 (10%). Accurate mass (M+) 200.0941 (required for C'2H'2N20 (M+)
200.0946 ).

[3H]pinoline. Characterization was determined by NMR and comparison with

cold pinoline produced under similar reaction conditions. Apparently all of
the 6-MeO-DHI3C was converted to product since none of the precursor could
be detected by NMR. Specific radioactivity (S.A.) of the product, as
determined by , Hand 3H-NMR, was determined to be 35.9 1.8 Cilmmol. The
NM R for the product was found to be in agreement with the spectrum of
unlabelled pinoline. The product was a pale-yellow powder.
360 l.C. Callaway er al.

','~,
H T
R= H,T

i
I
I
I
If.t
. ~
0 ..
~~
I\.,; .Ai

~ ....

I r , t' " I I I r
7~O
I 'i f 'f j i
9 .0 8.0 6'0 5.0 4.0 l.O 2.0 1.0

pp m

Figure 3 . Proton spectrum of [lHJpinOline in CD)OD ; note the

integral values for the doublet at 4.39 ppm resulting from tritium
and hydrogen atoms attached to Cl .

Table 1. 1 H-NMR of [3H]-Pinoline:

Atom ppm H, mult.

1 4.39, d
3 3.05, m
4 3.52, m
5 6.97, d
6- 0CH3 3.81, s
7 6.80, dd
8 7.23, d
Synthesis of ['H] Pinoline 361

Specific radioactivity. Measurements are generally made by liquid

scintillation counting of a known mass of compound, by mass spectrometry,
or by consideration of the 3H and 1H NMR spectra (12). The NMR technique has
been used to analyze compounds such as tritiated NaBH4, where other
methods are not facile or practical (13), and is most readily applied when
the amount of 1H replaced by 3H at specific positions is high; i.e. 10-100%
(14) .

One NMR method for determining S.A. involves the integration of

residual protons at the tritiated site relative to an unlabelled position with
a known number of protons, and this requires only a proton spectrum of the
product. Several issues need to be addressed when using this procedure: the
substrate must be chemically and radiochemically pure; the residual protons
are assumed to have the same T1 as the integral reference protons; the
proton spectra must be sufficiently well resolved to allow integration of
the sites of interest; and the SA of a substrate containing one tritium atom
per molecule is assumed to be 28.72 Cilmmole. In general, we consider that
a conservative (5-10%) error in NMR estimations of the S.A. should be
assumed.
In the present example a different approach was taken, utilizing both
the tritium and proton spectra. The same care with assumptions should be
applied as described in the first NMR method for S.A. assessment. In
compounds where multiple tritium atoms may be incorporated on one carbon,
the resonances of the isotopomers are resolved because of the primary
isotope effect on the chemical shift. This is true for both proton and tritium
spectra (Figures 3 and 4 respectively) , and comparison of the two reveals
the presence of the three possible species: R-CT2, R-CTH, R-CH2 . Note that
the R-CTH isotopomer has both tritium and proton spectra. This occurrence
allows us to assess the mole fractions of the three species in the mixture,
and thereby calculate a specific radioactivity for the product. Note also that
the intensity (or integral) of each tritium signal is proportional to the
number of tritium atoms observed, and not to the mole fraction of the
isotopomer (i.e. 0.5 x R-CT2 integral should be compared with R-CTH). Once
362 l .C. Cal/away er al.

the mole fractions of the three isotopomers (Mn,MTH and MHH) are known, the
S.A. is readily calculated:

e.g. S.A. - (57.44 x Mn) + (28.72 x MTH) + (0 x MHH).

8 .0
ppm

FIgure 4. Protoncoupled tritium spectrum of (3HJpinoline: the

larger peak represents RCT2 while the smaller partially
represents RCTH. the other half of the doublet is occluded by the
larger peak .

In our example, integration of the proton-decoupled tritium spectrum

(Fig. 5) easily yields the comparative proportions of the R-CT2 and R-CTH
species. In the proton spectrum there is overlap of the R-CH2 singlet and one
line of the R-CTH doublet, and this requires calculation of the R-CH2 peak
area (or accumulation of a tritium-decoupled proton spectrum). Combining
these pieces of information gives Mn .. 0.37, MTH - 0.51, and MHH ,. 0.12. This
yields a calculated S.A. of 35 .91.8 Cilmmole for the product.
Synthesis of [JH] Pinoline 363

y y.
M
"'N,
N
..
~
M
..,

Figure 5 . Proton-decoup~ed tritium spectrum ; selectively

decoupled at the signal resulting from the proton on C-l.

RESULTS AND DISCUSSION

[3H]pinoline has been synthesized with high specific activity by

reducing the hydrochloride salt of 6-MeO-OHI3C with tritium in the presence
of a catalyst. The tritium form of this endogenous compound was prepared as
a chemical probe for binding sites associated with the serotonergic system_
Pinoline has been shown to displace specific ligands of the 5-HT transporter
complex in nM concentrations (3). We hope to learn more about its
endogenous role and putative binding site(s) by using (3H]pinoline as a
radioligand_ In addition, with the development of a method for producing
stable hydrochloride salts of OH I3Cs which are unsubstituted at C-1, we can
expand our investigations into this new group of I3Cs.

Acknowledgements. Jukka Knuttinen and Seppo Auriola for their MS

expertise. A grant from the Finnish Academy of Sciences and Finnish
Cultural Fund. A grant from NIH, P 41-RR01237, which supports the National
Tritium Labeling Facility.
364 l.C. Cal/away er al.

REFERENCES

1. Adichi J., Mizoi Y., Ogawa Y., Uetani Y. and Ninomiya I. - Journal of
Nutrotion 12.1: 646 (1991)

2. Adachi J., Yamamoto K., Ogawa Y., Ueno Y., Mizio Y., Tatsuno Y. - Archives
of Toxicology.fi5.: 505 (1991)

3. Airaksinen M.M., Gynther J., Po so A., Callaway J.C. and Navajas C. - British
Journal of Pharmacology .1..Q1: 370P (1991)

4. Airaksinen M. M. and Kari I. - Medical Biology 5.9.: 21 (1981)

5. Buckholtz N. S. and Boggan W. O. - Biochem . Pharmacol. 26.:1991 (1977)

6. Fryer R. I., Cook C., Gilman N. W. and Walser A. - Life Sciences ll:1947
(1986)

7. Gynther J., Lapinjoki S., Airaksinen M.M. and Peura P. - Biochem . Pharmacol.
~: 16 2671 (1986)

8. Ho B. T. Current Developments in Psychopharmacology 1... Spectrum

Publications, Inc. (1977)

9. Maclaren J. A. - Aust. J. Chem ~ 1617 (1987).

10. Nimit Y., Schulze I., Cashaw J. L., Ruchirawat S. and Davis V. E. - Progress
in Clinical and Biological Research a.o..... Beta Carbo lines and
Tetrahydroisoquinolines, Alan R. Liss Inc., N.Y., p. 311 (1982)
,
11. Pentikainen H. T., Airaksinen M. M., Tuomisto L. and Peura P. - Alcohol and
Alcoholism 2.:1, 33 (1986)

12. Evans E. A., Warrell D. C., Elvidge J. A., Jones J. R. - Handbook of Tritium
NMR Spectroscopy and Applications, Wiley: Chichester (1985)

13. Altman L. J., Thomas l. - Anal. Chem. ~, 992 (1980)

14. Bloxsidge J. P., Elvidge J. A., Jones J. R., Evans E. A., Kitcher J. P., Warrell
D. C. - Org. Mag. Reson. 15., 214 (1981)
III

o 1993 Elsevier Science Publishers B.V. All rights reserved.

Melatonin and the pineal gland - From basic science to clinical application

Y. Touitou. J. Arendt and P. pevet. editors. 83

Binding sites for [3H]pinoline

M.M. Airaksinen l , I.C. Callaway l.2, P. Nykvist l , L. Rago l , E. KariI and I . Gyniher2

1Departtnents of Pharmacology and Toxicology and 2Pharmaceutical Chemistty, University of

Kuopio, P.O. Box 1627, SF-70211 Kuopio, Finland

INTRODUCTION
Pinoline (6-methoxy-I,2,3,4-tetrahydro-B-carboline, 6-MeOTI-lBC, 5-methoxy
tryptoline) has been found in similar concentrations to those of melatonin in mammalian and
avian pineal [I] and retina [2], where its endogenous synthesis can proceed as shown below.
The cyclization of serotonin (5-HT) to 6-hydroxy-TI-l13C (6-0H-TI-l13C) and subsequent
methylation may be the main synthetic pathway, though pinoline may be formed non
enzymatically from 5-MeO-tryptamine (5-MeOT). The cyclization oCthe tryptamine sidechain
occurs through a non-enzymatic Pictet-Spengler cyclization using glyoxylic acid followed by
an enzymatic decarboxylation of the terrahydro-13-carboline [3]. Pinoline's physiological
function is not known, but it inhibits 5-HT uptake, is taken up into synaptosomes and inhibits
MAO-A in 5-HT neurons. Tritiated pinoline (l,I_[3H]) was synthesized in order to investigate
potential binding sites in rodent and bovine pineal glands, brain and adrenal tissues.This
binding was displaced by B-carbolines found endogenously but only partially by specific 5
HT uptake inhibitors.

HOQD
H
----_r ~ ~OMT
5-HT
6-0H-THBC ~c~
VZ.. ~~
N

/ H
pinoline
(6-MeO-TH BC)

melatonin 5-MeOT

MATERIALS AND METHODS

Adult male Wistar rats (300-320 g, National Laboratory Animal Center, Kuopio,
Finland) were kept under standard laboratory conditions with lights on from 07:00 to 19:00.
Tissues were collected during midday. Bovine pineals were obtained from a local slaughter
84

house, also during the day, these animals were exposed to natural and artificial light prior to
slaughter. All tissues were collected in October.
After removal, tissues were immediately frozen on dry ice (in isopentane for
autoradiography). Tissue sections 14 mm thick were thaw-mounted on gelatin-coated glass
slides, and dried under vacuwn at 4 C. Tissue sections were washed and labelled with 10 nM
of [3H]pinoline (with and without 1 mM of unlabelled displacer) at 4 C for 60 min ., rinsed
and dried under air. Slices were exposed to [3HJHyperfilm (Amersham) for 8 weeks.
Autoradiograms were developed by using Ilford PQ Universal Developer. Binding studies
were performed as previously described [4].
Pi noli ne, tetrahydro-B-carboline (TI-lBC), 5-HT(creatinine sulphate), melatonin and
pargyline were purchased from Sigma. [3H]Pinoline (spec. act. 36 Ci/mmol, radiochemical
purity> 99%) was synthesized as previously described [5].

RESULTS

The highest density of binding sites were associated with the pineal gland in both
species studied, as determined by autoradiographic techniques. (Fig. 1 A & B, Fig. 2 C &D).
High densities were also located in rat adrenals, both in cortex and medulla (Fig . 1 C & D),
with somewhat lower densities found in rat brain (Fig. 2) and bovine cerebral cortex (not
shown). The binding distribution was non homogeneous in the brain , with higher densities
seen in the gray matter of cerebral and cerebellar cortex. Particularly high densities were also
observed in the interpeduncular nucleus, median raphe nuclei, septal, hypothalamic, pontine
and central periaqueductal grey areas (Fig. 2).

A B

Figure 1. Binding of [3HJpinoline in the pineal gland of cow (A, B) and in the adrenal gland
of rat (C, D,E). Total binding (A, C) and nonspecific binding after displacement with
unlabelled pinoline (B, D) and pargyline (E).
 85

In the rat brain, specific binding of [3H]pinoline could be displaced by the following;
TH13C ~ pinoline > S-HT ~ melatonin> pargyline ~ citalopram = paroxetine by using 0.1-1
mM of the displacer. Only partial displacement of [3H]pinoline was achieved with the S-HT
uptake inhibitors paroxetine and citalopram, even at high concentrations. The MAO inhibitor,
pargyline, also accounted for only partial displacement except in adrenal medulla, where it
effectively displaced [3H]pinoline.
Of all the displacers studied, TIffiC had the highest ability to inhibit binding in the pineals
of both species. The ICSO values for TIffiC in cortex and pineal membranes were found to be
112 nM and 74 nM, respectively.

Figure 2. Binding of [3H]pinoline in rat brain, 11 mm (A, B) and 2 mm (C, D) rostral from
interaural level. Total binding (A, C) and nonspecific binding after displacement with
unlabelled pinoli ne (B) or TIffiC (0)

DISCUSSION AND CONCLUSIONS

Apparently [3H]pinoline does have specific and high affinity recognition sites in
adrenals, pineal and brain tissues. Pinoline has high affinity for the 5-HT transporter and
displaces [3H]imipramine [6,7], [3H]paroxetine and [3H]citalopram [8, and our unpublished
results] from their recognition sites, however its binding distribution in brain only partially
resembles the autoradiographic data obtained from 5-HT uptake inhibitors [9]. Moreover these
antidepressants only partially displaced [3H]pinoline from its binding site, suggesting that
these are not the only binding sites for pinoline. Saturation studies also indicated more than
one binding site (data not shown). Part of these binding sites are probably within MAO-A,
while a portion seems to remain distinct from it and the S-HT transporter. Therefore pinoline,
as an endogenous 6-carboline, may modulate the serotonergic system by increasing the local

,
86

concentration of unconjugated 5-HT, and might also act through its unique and specific
binding sites.
High and specific affinity for pinoline, together with its production and significant
concentration in the pineal gland, indicate its local function in that organ where it can influence
melatonin synthesis by inhibiting serotonin's uptake and oxidation. It also had high affinity
for adrenal tissues. In adrenal medulla the binding site of pinoline may simply be MAO; in
adrenal cortex the nature of its binding remains to be investigated. This high affinity in
adrenals reopens an old discussion on the pineal's influence on aldosterone secretion [10, 11]

REFERENCES

Kari I, Airaksinen MM, Gynther J, Huhtikangas A. Rec Devel Mass Spectr Biochem Med
Env Res 1983; 8: 19-24.
2 Leino M, Kari I, Airaksinen MM, Gynther J. Exp Eye Res 1983; 36: 135-138.
3 Gynther J, Lappinjoki SP, Airaksinen MM, Peura P. Biochem Phannacol 1986; 35:
2671-2675.
4 Kari E, Tuomisto L, Airaksinen MM. Exp Eye Res 1988; 47: 679-688.
5 Callaway JC, Marimoto H, Gynther J, Airaksinen MM, Williams PG. J Labelled Compds
Radiopharm 1992; 31: 355-364.
6 Langer SZ, Lee CR, Segonzac A, Tateishi T, Esnaud H, Schoemaker H, Win bland B.
Eur J Phannacol 1984; 102: 379-380.
7 Airaksinen MM, Kari E. Adv Biosci 1991; 82: 239-240.
8 Airaksinen MM, Gynther J, Poso A, Callaway JC, Navajas C. Br J Pharmacoll991; 104:
370p.
9 Hrdina PD, Foy B, Hepner A, Summers R. J Phamacol Exp Ther 1990; 252: 410-418.
10 Farrell G, Mclsaac WM. Arch Biochem Biophys 1961; 94: 543-544.
11 Airaksinen MM, Sainio E-L, LeppaIuoto J, Kari I. Acta Endocrinol 1984; 107: 525-530.

This study has been supponed by the Academy of Finland (Medical Research Council) and the
Finnish Cultural Foundation.
IV

 MarApr 1994 The Pictet-Spengler Reaction and Biogenic Tryptamines: Formation 431
of Tetrahydro-~-carbolines at Physiological pH
James C. Callaway' [a,bl, Jukka Gynther [a), Antti Poso [aI,
Jouko VepsliHtinen [cl and Mauno M. Airaksinen [bl

Departmenu of Pharmaceutical o,emislry, [01

Pharmacology.od Toxicology, [bl .Dd o,emislry. [cl

UDiversilY of Kuopio P.O. Bo. 1627,

F1N70211 Kuopio. Finl... d

Receive<l Novembec S, t993

Biogenic tryptamines la-c were reacted with aldehydes 2a & b and akelo acids 2e & d la form
1.2.3.4.tetrahydro-p-carbotines (TIlDes) 4dt. and other products. in a buffered solution at 37 and pH
7.4. These reactions were followed over time by III runr through integral changes in discrete signals in the
spectra. Reaclions belween tryptlllllines and acetaldehyde (2b) gave.lhe eapected lmethylTIIDCs 4dr,
while those with sodiwn glyoaylate (2e) resulted in TllDCIcarboaylic acids 41:'1. Surprisingly. reactions
with sodiwn pyruvate (2d) or formaldehyde (2a) diu not form the eapecled products 4ae or 4JI. respec
tively under these conditions. In successful reactions. 5methoaytryptamine (le) was found to be more
reactive than tryptamine (la) or serotonin (I b). MOrAC calculations were employed to invcstigate rcac
tion intermediates . These results arc applicable in research related la aberrwlltryptamine metabolism ; . g.
depression and alcoholism.

/. Hew"cyclic Ch..... 3l 431 (I994~

~-Carbolines (9H-pyrido[3,4-blindoles) are considered
to be natural components of many plants and animals [11
and display a broad range of biological activity [2).
Recent evidence continues to support the idea that
1.2.3,4-tetrahydro-p-carbolines (later on, TImcs) are rou
tinely formed in humans [3). which encourages specula
tion on their putative role in Llle central ncrvous system
(CNS). Some of Llle THDCs derived from tryptamine (la)
and 5-methoxytryptamine (le) inhibit monoamine oxi
dase-A [4]. and bind wiLll nM afflllity in Llle CNS to Llle
serotonin transporter [5.6,71. Thus, through entirely dif
ferent mechanisms, THBCs are capable of modulating
serotonergic activity by preventing the metabolism of
serotonin (lb) and its ncuronal rcuptake, respectively.
This is important since abnormalities in serotoncrgic
activity have been implicated in a host of neurologic dis
orders, including psychiatric illnesses such as depression
[8,9i, neurodegenerative diseases like Alzheimer's and
Parkinson's diseases [IDI, and alcoholism [11). Also, p
carbolines have becn implicated, at least to some extent.,
in all of Lllese disorders [12,13,141. As hypoLllesizcd earli
cr, this may be especially relevant in Llle casc of alco
holism, where abundant amounts of acetaldehyde (2b) are
available to react wiLll tryptamines 1a-e to fonn the I
meLllyl-TI-lDCs 4d-f (later on, I-Me-TIlDCs) [15,16).
The tryptamines la-e are readily found in humans.
Small amounts are taken in directly through the diet,
while most are derived from the esscntial dietary amino
acid tryptophan. Conventional wisdom suggests that
endogenous THDCs are formed nonenzymatically
through a Pictet-Spengler condensation between these
tryptamines and carbonyl substrates [171 (see SCheme),
similar to Llle formation of tetrahydroisoquinolines from
pheneLllylamines [181. This reaction is especially favor
able towards pheneLllylamines under acidic conditions.
and speculations on Llle endogenous formation of TllDCs
from tryrtamines arc routinely extrapolated from Lllese
results, UlOugh rcports of careful investigations into simi
lar reactions wiLll uyptamines at physiological pH (7.4)
are lacking . In our continuing efforts to investigate Llle
ocurrence and actions of endogenous TI-lDCs, we decided
to examine more c10sel y Llle reaction deemed responsible
for their biological ocurrence; i.e. the Pictet-Spengler
reaction. Reported are Llle results of individual reactions
between tryptamines 1a-e with two common aldehydes
(formaldehyde 2a and acetaldehyde 2b) and two sodium
salts of et-keto acids (gl yoxylale 2e and pyruvate 2d) as
likcly biological carbonyl sources. AI ~o reported. and dis
cussed. are Llle results of MOPAC [19) point charge calcu
lations of the carbon acorns connected prior to cyclization.
Results and Discussion
Adduct formation is presented graphically for Llle for
mation of Llle THDC-I-carboxylic acids 4g-i (later on, I
COOIl-THDCs) from 2e and 1a-e (Figure 1), and Ule for
mation of I-Me-llmCs 4d-f from 2b and 1a-e (Figure 2),
over time. Overall reactions wiLll 2e as Llle substrate were
initiated faster (as determined by Llle y-intercept) and pro
ceeded at a faster rate, than reactions with 2b, and in both
cases le was found to be more reactive Lllan eiLller la or
Ib (Table I). Tryptophan was not sufficiently soluble
under Lllese conditions and excluded on this basis.
 432 J. C. Callaway, 1. GynLher, A. Poso, J. Vepsillliinen and M. M. Airaksinen Vo!. 31

Schema Table I

Resulu of IH NMR Experimcn u of Tryptamine.s (1a--<:) Reacted wilh

R Glyoxylale (2e) and AcclaJdchyde (Zb). All Rea<:t..ionJ were Run in

~NlI, Triplic~e .
I.W

~))l +
N
2. HP nM rc lcled nM/min
H
(y ilIlQ"c c-p4 ) ( I lop")
"
Glyoxylale reacti ons
I .., l.-d J .-1 Tryplamioe ICooH TIIOC

! H'
la
Ib
le
4g
4h
4;
D8nM
J.25
4.46
0.1 J3
0.175
0.332
0.998
0981
0.993

0:0,11
R'

~ RI H
~
R'

~~'II
N RI R,
AceuJdch yde reaction,
Trypuminulee
I.
IMe- TIIDC
4d 0.88 1 nM 0.067 0.994
Ib 4c 1.014 0.081 0.960
5.-r 4.1 le 4r 1.85 I 0.149 0.982

AD ioclination lowAnis form.ation o f nmes iJ inwc31ed by the y.intcr.

COlllround R RI R, Common n;unes
cepl. in nmole.!. TIle cales were: determined dHcctly (rom the slopc .~
I> 11 uypcam.ine (nMlminutes),o( lflpilcalC: runs . RC:3.cLJons with glyo~yble 2e were: well
11. 011 serotonin. S-UT under wJoy by t..hc lJme of lhe: first SPCClIJ. and co ntInued 10 procccJ :1.1 a
le OMe 5 MeO-lryJ>Camioe fasler rale: Ihan reactIons wilh aCClaJJchyJe (2b) . Of .IIJ the lrypIJrrunc:s.
b H H focmaldehyde 5mc:t.hoxytryplanune (le) wcu found 10 f caC1 moce rcawly lhan Itu: olh
Me H ers foe both carbonyl subSLralc:s.
2b aceuldehyde
2e H COO- glyoxylate (N.)
2d Mo COO- py",v..e (N.) from the III nnu speClIa. where a clear singlet for a pro
3,4,5. H H H nlDc ton on C-2 appears in the aromatic region (e.g., S 7. 11. s.
3.4,5b OH H H 6-110-THDC
3.4.5c OMe H H 6-MeO- TIIDC
III for the prooucI from la). Ullinlate characterization of
3,4,5d H Me H I-MeTIIDC these proouclS lies beyon\! Ihe scope of tJlis paper, as il is
3,4.5. 011 Me H I -Me6-1I0-TIIDC evi\!cnt Ihat TllllCs are nol rea\!il y forme\! un\!cr Illcsc
3.4.5r OMe Me H IMe-6-MeO-TIIDC con\!itions by this reaction pathw ay.
3.4g H H COOl! ICOOIITIIDC
3.41. OH H CooH ICOOH6-HO-TI mc
3,4i
3,4J
OMe
H
H
Me
CooH
CooH
ICooH6-MeO- rusc
IMeICooHTIlDC
'.0,--------------------------------------,
3,4k OH Me CooH IMeICooH6-HO- ruBC
3,41 OM. Me CooH IMe-ICooH6-MeO-THBC 120

TIle Piclet-Spengler rC.lctioD in the condensation of tryptamiDes la-<

wilh aldehyde.< h & b or ,!<clO . . id, 2e & d 10 form TIlDe.. 4 ..,. 1
...
MeTHDe.. 4dr. 'nd ICooH-TIIDe.. 4gllhrough a cationic inlermc
di .lle Ja-I which follows the formJ.lion of a Shire base (no l shown). Also
j 100

show n are lhe TI-IDCs obl3ined after acid hydrolysis of l -eaOH U

80
TI -t oCs. 53'(, derived from the aketo aciw. The commoo names listed '"
do not ::apply (0 inlermwia.le..!. ':'
::
0
0 60
'-!
f'avor ab lc reactions with 2d to form 4j-1 were nOI
ohserved under these conditions. The pyruvale reactions .. .0

onen resulted in clear dark-red solutions of complex com ~

position after several days al 37. 20
Reaclions between 2a and la-c consistently and inune (4,) l e aQ HTlIDe
\! i;lIdy resu lle\! in the precipitation of complex pnxlucts
which were not the expecled TIIDCs 4a-c. Instcad. these 10 20 '0 .0 50 60 70
pro\!uel s more resembled tryplamines with repealing
~cq uences of -CHrO- all3ched to the aliphatic nilIogen Time (mlnulu)

(f'igure 3). f'airly pure samples of these products were

Figure J. f'ormalion of l-COOII-TIfIlCs 4g-i over lime. in nmoles.
oblained when this reaction was run for 1 minute at 0, (rom reaclions between tryptamines la-c and glyoxyl:lIe (2t:) .
and none of the products had properties consislenl with Precipil.3UOn of Ihe proJuCI prevenlccJ 3quisilion of suilabl e spectra from
compounds 4a-c. The prominent evidence for this comes 4i and 4g ailer 25 anc.J 40 rninulcs, respectively.
Mar.Apr 1994 Formation of Tetrahydro-~carbolines al Pbysiological pH 433

..
~
140

120 -- (41)

(4
1M.6-MeOTHBe
,M..6HQ.THBC

~ 100

..=....
U
80
R. H. 011. 0C11)

~ 60 Figure 3. Polymeric N melhyJohrypCarrune.r resuhed from reacljons

. 40
~
20
to 20 30 40
TIr... (,.Inuln)
Figure 2. Formation 0( JMe-TJ me.:
4d' over Lime. ia nmoles.
rC.Jcuons between tryplamiac.s la-c and iIIcclillldchyck (lb).
Half of our expcrirnenlal protocol resullcd in unexpect
ed produCIS, since we expecled la-e 10 react with both
aldehydes and both aketo acids 10 form 11113Cs, as pre
dicled from the PicletSpengler reaclion. To funher exam
ine this anomaly, semiempirical calculalions (AMI) were
inilially performed 10 invesligale why 2b readily reaclS
with lryplamines to form THI3Cs while 2a does not, and
ultimately to examine the electronic propenies of other
adducts (Table 2). The results of these calculalions
revealed several interesting features. For example, Ihe
indole C2 position on the intermediate 3d carried a nega
tive charge (0.064) while the carbocalion had a posilive
charge (+0.135). Such a difference in charge is apparently
suflicienl for cyclization, similar resullS were obtained for
:le & f (Table 2). When intermediate 33 was calculaled,
tile charge on C-2 was found to be -0.036 with a calionic
charge of +0.063 . These values for 3a were almosl half
those rcsulling from calculations for 3d. Similar valucs
were oblained for intermediate 3c, and even 3b where the
negati ve point charge on C2 was quite large (-0 .051) ,
ulough apparently not large enough to induce cyclizalion.
belw~D ltypIamiD'" hc aDd fO"'lAhJehyde (Zo), where X inCJe=~
with time .Dd lempcrllure.
For the sake of comparison, similar calculations were
made for phenethylamine (later on. PEA), since this corn
60 60 70
pound is reponed to react with both aldehydes to form
tetrahydroisoquinolines [17,18}. The calionic point charge
for the intermcdiate calion of PEA with 2a (Table 3) was
from
not found to be much differenl than those for 3a-e (Table
2). However. the crucial points were found to be at the
two possible sites for cyclization on the aromatic ring of
PEA. onho to the ethylamine side chain. where presum
ably the reaction lakcS place at the most negative site.
Here. the more negalive of the two point charges for ule
intermediate of PEA. with la as the substrate, was found
to be 0. 160, and with 2h as the substrale 0.148. This is
roughly four times more negalive than C2 posilions on
3a-e, while C2 on inlermediales Jdf was only about 2.5
limes more negative. In addition, the absolule diffcrential
in charge between the calion and C2 was quile large for
intermediates 3d-f, relalive to 3ae.
With 2e as the substrate, the more negative of the two
ortho positions on PEA was 0.123, which was almost
twice as negative as the charges on C2 for 3gi, The
effect of 2e on the positive characler of the cationic
charge helps explain ils rcactivity towards the aryl carbon
since the global differenlials. that is the overall difference
in charge bctween the calion and C2. were all negalive
for 3g.i and inlermediales of PEA . Thai our calculalions
with PEA always resulied in a global negative difference
between the point charges on the ortho posilions and the
cation. and that the absolute dilference was relalively

T,bl. 2

MOPAC Calcui>tioM 01 Poinl o.uges (cation an~ C2) on Tryptamine< la.., .n~ Intermediate. (int.) 301,

aoolhe absolu te diffcrentiab (... w. dieL) between these valuc.s.

lnlcrmecliale.s of la InlermecJ ialcl of Ib Intermediates of le

charges hs. chorS'" ,b.s . ctu.tge.s ab.s.
into c300n C2 ~ill. int. caLion C2 ~ il r. inl. c3tion C2 dill.

3:.1 0.063 0.036 0.099 3b 0.073 O. O~I 0.124 3e 0062 0.030 0.091
;Id 0.135 0.064 0. 199 l< 0.135 0.056 0.191 3r 0.136 0.056 0.191
3g 0.011 0.074 0.085 3h 0.013 0.065 0.078 3i 0.013 0.066 0.079
3j 0.054 0.071 0. 125 3k 0.048 0.049 0.097 3t 0.054 0.062 0.116
Iu 0.081 Ib 0.073 le 0.072
Values obtained from poinl charge catcuJaJ.ions (MOPAC 5.0) of the carbon atoms connected just 10 cycli7AI.uon; the C-2 carbon on the indole ring. and
UIC intermediate carbocation aluched to !.he aJjphatic nitrogen of the inlerm<.-di3le 3a-l, ano U1C point charge.s for C-2 on the unrcactcd IQ lIypl31'1'Unes
hoc. Also shown are the absolute differences ia charge: between tIle Calion ano C-2.
434 1. C. Callaway, J. Gyntber, A. Poso, 1. VepsaHiinen and M. M. Airaksinen
Table 3
MOPAC Clllcul.1tioD.! of Point Otarge.s (catiooic And uyl posiuoQ.t) few
intriguing in the case of excessive ethanol consumption,
PEA and iu inlcrmedialc.s. Differentia1.t were CalClJlaled (rom the MOfC
since approximately 0.9 g of 2b can result from every
Negative Aryl Value
di~~j&1
catioruc charge OD absolute
subsu;uc: charge onho positiolll differe.tiaJ
2a 0,062 -0, I 60. -0, "2 0,222 -0.098
2b 0,124 -0. I 48. -0, "4 0,272 -0.024
2c 0,006 -0, I 23. -0,123 0,129 -0,117
2d 0,047 .0,124 . .0,122 0,171 -0,077
1 J.mine 0,138. -0. I 26
V,lue.s obuined from poinl charge calculatiOn> (MOPAC 5.0) 0( Ihc car
bon &EarN connected pria to cycliz.atioD for pheDelhylaminc (PEA); the
IWO onlw pol iuons on the &lomllouc ring. ADd Lhe intermediate C&lboca
Lion on Lhe ~liph;&t.ic wtrOgCD . and tjlC poinl ChMge3 on the: atOrN onJro 10
the eLhyl.mu nc side d13jn of unre.3.cled PEA. The ~olule and glob.1l dif
ferentials were caJculaled (rom !.he point <:huge value ollhe ll'lOSt
live OrfJro ca.rboa.
large, is consistent with earlier observations showing that
this reaction clearly favors the cycli7.ation of PEAs over
u'ypramines [20 and references therein].
The unreactivity seen with 2d presents a special case,
and our observations arc in agreement with earlier work
['ubli s hed which demonstrated that 2d does not readily
condense with la betweenpI17.1-7.4 [2IJ, Unlike 2e. 2<1
lacks a prolon on ule p carbon and must form an enol
before addition of the amine to the subsequent double
bond [reference 13 in 17J. And unlike reactions with 23,
2<1 does react with tryptamines under weakly acidic con
ditions and readily decarboxylates to 1-Me-THllCs
nonenzymatically [22].
The polymeric products (Figure 3) from reactions
belween 2a and la-c were unexpected. Apparently the
reaction would require a much lower pH to form the
iminium Shiff base, which is essential for cyclization
[23.24]. and unlike 2b, 2a is exceptional in that it lacks tl
hydrogcns on the carbon adjacent to the carbonyl and lhe
illlcrmcdiale can nOI be stabilized through hypcrconjuga
lion [25J , Also. with 2a as the substrate. the native point
charge on C-2 may nOI be sumciently negative for trypta
mines lac to attract the intermediate cations of 3a-c into
seeking stability lO form TImCs 4a-c through elec
trophilic cyclization . Instead, the intermediate cations
seek additional formyl carbons under these conditions.
Thus, when making the common assumption that
THllCs can be made from tryptamines and aldehydes, one
should bear in mind the reaction conditions. particularly
pi I. This same tendency of tryptamines to react with 2a
was a major obstacle to ule initial verification and accep
tance of THllC production in vivo, since trace amounts of
2a can be generate d by biological samples during the
work up and typically persists in many extraction sol
vcnts. also these extraction procedures were inevitably
undcr very acidic conditions [26J.
Vol. 31
The results from reactions with 2b are especially
gram of ethanol conswned. It has been shown that acute
and chronic increases in 2b result in the endogenous for
mation of 1-Me-THllCs in humans and other mammals
[14,20,21,27-30J, and an acetaldehyde-induced imbalance
between 1-Me-THllCs and I-II-THIlCs seems reason
able, This hypothesis is also encouraged by the reactivity
seen with ~taJdehyde (2b), relative to glyoxylale 2e in
regard to these trypt.amines I a-e, and by the fact that the
pharmacology of I-Me-TIIDCs and I-H-TllDCs differ
marl<edly [3-14J.
nep EXPERIMENTAL
An.lyses of Ihe re.elions were m.de by III and IlC nmr.
spectra were recorded over lime on 8 Druker AM 4001 AS[>CCl
3000 nmr s[>CclIomcler o[>Cr>l ing >I 400 .134 Milt for II1 'fltJ
100 ,614 Milt for IlC S[>CCII>, The s[>CCIt, were .cqu ired u' lOg
32 kW d.1.:! poinlS wilh tero filling 10 poinl resolulion beller
Ihan 0,2 IIz, Idcnlilics of all final products were fu"her con
firmed by El mass spcc(romctry. recorded on a Trio-2-VG
Massl:lb spcclromclcr (Mam.:hcslcr U.K .) by din': Cl inJeclion
probe Jl 70 cV Wilh an ion source Icmpcr:uUfc of 200 anti an
clcclIon current of 200 ~A. ElcmcntaJ compositions of lhe iuns
were confirmed by accurate mass measurements: and were in
.grccmenl wilb previous work I3t),
For the lime course reactions, cquimolar concentrati o ns of
Iryptamine (ltypl.1mine hydrochloride and 5melboxylryplamine
hydrochloride, Sigma; or serolonin crealinine sulph.le. Merck)
and carbonyl subslr:lle (formaldehyde 37% aqueous wilh 10%
melhanol. ,ccl:udehyde 99%, Merck; sodiwn glyoxylale, nuka;
or sodium pyruv.le. Doehringer). were reacled in 5 mm nmr
lube; 0 .64 mmoles of Ihe dissolved Irypl>mine (as a 400 III
:uiquollaken from. 1.6 M aqucous solulion. pll 7.4) was added
10 700 III of buffer (0.1 M aqueous sooium phospale. pll 7.4). fol
lowed by 100 III of TSP (sodium-3-(ltimelllylsilyl)-2.2.3 .3[d 4 )
propionale. Merck) and 100 III deuleriwn oxide (Merck), l'inally.
0.64 mmoles of Ihe carbonyl subsltale was added >s a 50 III
aJiquol laken from a 12,8 M aqueous solulion , lbe runt lube was
brieny shaken (I =0 minules) 10 mix Ille reaclants prior 10 place
menl inlo Ibe probe (al 37), Afler shinuning Ihe insltumenl. the
inili:u IJI spec Ita were oblainable ailcr 3 minules , Dal:! from Ibe
integr.led speclt. of Illese re.clions were plo lied as a nonn.lized
percenl.ge of lhe producl formed. expressed in nmoles. versus
lime. in minu\cs, The nmolar amounl ofTIIDC fonned .nd Itypl
amine reacled. were caiculaled directly from Ibc inlegr:u value of
single prolan signals in the arom:.uie regio n of the spectra .
Preeipil'lion of lhe prooucl lypically marked Ihe end of suc
cessful reaclion. :is well as Ibc al>ilily 10 oblain gooo 'peclt . All
reaclions wcre run in Itiplicale. Acelaldehyde was diSlilled prior
to use.
Addilion:u IH nmr analyses were made of prooucts resulling
from reaclions wilh Ibe a-keto .cids 2c & d, following dec,,
hoxyl.lion of Ibe rcsulling I-COOII-TIIDCs 41:.110 Ibeir corre
sponding l-ll-TllDCs 5a-f by the addilion of lOO III 37%
MarApr 1994 Formation of TetIahydrl>-~-carbolines at Physiological pH
hydroc hl o ric acid.
For anal ytical purpos 0.100 g (0.51 mmol.) of la
hydrochloric acid was reacted with an equimolar amount of la
for one minute at aa, since bigber temperatures and longer reac
tion times resulted in compl.x resins. A fin.white electrOSlatic
powder was recov...ed (0.107 g) after nItration and desiCGation,
which foam.d and decomposed ov.r a wide temperature range
(85.125). This foam was. red oil by 165 and a clear darlc red
oil by 290. The product did not dissolv. in common organic
solvents, and decomposed after the addition of organic and min
eral acids. It was insolubl. in 5 M sodium hydroxide and con
centrated ammonium hydroxide. tH MU (DMSO d 6 ): [,2 .8 1
(br. Ill, N H), 2 .842.87 (m, .thyl, 4H), 3.41 (s, methylen IUld P. Bouely.l-</eSci.. 48,1977 (1991).
6H). 3.51 (br, IH , OH), 6.96 (dd, IH, 5H, J = 7.0, 7.8 Hz), 7 .05
(dd. Ill. 6H. J ~ 7. 1. 8.0 Hz). 7.11 ('. IH. 2 H). 7 .32 (d, IH, 7
11. J = 8 . 1 IIz), 7.52 (d. IH. 4H. J = 7. llIz). 10.71 (s. IH. ArN
11): IlC nmr (DMSOd 6 ): [, 136 .2. 127 .2. 122 .4. 120.7. 118.3,
118 . 1. 112 .5. 111.3 , 73.7.52 .9 , 51.8.23.3: m.: mJz (relativ.
ifHensities) 186 .09 (9), 172.14 (23).143 .73 (base). 129 .95 (25),
115 .22 (6). Small amounts of impurities w ..... vident from th.
nrnr spec tra and all attempts to purify this product resulted in
more complex mixtures.
Except for substi tutions on th. aromaLic ring, th. spectral and
physical properLies of analogous products resulLing from I band
I c were essentiaUy id.nLical and otherwise unremarkabl. (data
no t shown).
Structures of the intermediate reaction products 3aJ were
fully opt imized using a semi . mpirical MOPAC 5 .0 program
with AM I param.terization [19J . Point charges were obtain.d
from standard output and compared with calcu laLions from anal
o go us intermediates of ph.nethylarnine (PEA).
Acknowl edgements.
This work was sup ported in part by grants from the Finnish
Academy o f Scie nce. Medical Reseorch Council. and Ihe
Finnish Culiural Fund. Thanks 10 Mr. Jukka KnuuLinen for mass
spectral analyses and Mr. Olavi Manninen for compuler mainle
nance.
REFERENCES AND NOru
I11 M. M. AiraksiDen aod I. Kari, Med. BioI. . 59. 21 (1981).
121 M. M. Airiliinen and I. Kati. Med. BioI. . 59.190 (1981).
131 I. A~aehi . Y. Mizui, T. N1i.o, Y. Ogawo. Y. Ue.ani, and I.
Ninomiy . J. Nu".. 112. 646 (1991).
14) N. S. Duekholu and W. O. Bo"an. Biochem. Pharmacol.
26.1991 (1977).
15) M. M. Airaksinen, J. C. Callaway, P. Nykvis~ L Rigo. and
1. Gynulcr. in Mel3lonin and the Pine.al Gland. r.,.orn
Basic Science 10
Clinical Application. Y. Toui(ou, J. Arendl and P. Ptvel eds. Elsevier
435
Seie""" PubliC&lioDS B. V., Arruterd&m. 1993. pp g386.
(61 M. M. Airaksi"eD, I. Gynther. A. Poso, I. C. Call away, and
C. Navaju. Br. 1. Pharmacol. 104. 370P (199 1).
(7) M. M. AiraksineD and E. Kari. Adv. Biosci.. 82. 239 (1991).
(g) H. Y. Meluer aDd M. T. Lowy. iD Psychopharmacology: The
TIUrd GcncralioD of Progress. H. Y. Mehzer cd. RaveD Press. New
YOfk. 1987, P 513. .
191 S.-L Brown and H, M. van Prug, ill The Role of Serotonin
iD Psychiatric Disorder!. BrunnerlMazel. New York. 1991. ,
1101 R. J. D'Amalo. R. M. Zweig. P. J. Whi.ehouse. G. L. Wcnl<,
H. S. Singer. R. Mlyeux, D. L Price and S. H. Snydet. Ann. Neurol., 22.
229 (1987).
Ill) M. Daousl, I. P. llluintre. D. Elnouf, E. Lcgrand. P. Drelon,
(12) I\. W. Prooer aDd D. M. Drown, T,.nds Neurosci., 11 .209
(1988).
113) B. Te.su. R. NaylOf. D. Coslall . P. lenDer, C. D. M.,,<len, J.
Pharm. Pharmacol .. 37. 679 (1985).
)l4) O. Deck , T. R. DosiD, A. Londman. and S. Dorg, Biochem.
Pharmacol.. 31. 2517 (1982).
(15) W. M. Mds..c and R. T. 11mi . Adv. Pharmac.. 6. 247
(1968).
1(6) ~ D. 1101 ma n. G. R. Elli"... K Faull . and I. D. Darch .... in
PJychophllmacology of Alcoho l. M . Sandler, ed, Raven PrcS!, New
York. 1980. pp 155169.
(171 W. H. Whaley anti T. R. GovindolCh:ui. in Organic RC.lCOOM.
VoI 6. R. Adanu. 11. A~kilU, A. M. Dla", an~ A. M. Cope. e~s. lolon
Wiley&'SoDS,NewYork, 195l,pp 151191.
1181 A. Pie.e. and T. Spcnglct. Ba.. 44. 2030 (1911).
I J9) MOPAC 5.0 is .. genenl pwposc. scmi-emperlcnl molecub.r
orbit.a.i package de.s igned (or !.he sludy o( chemiCAl llructures and reac
tions which wa.s wrillen by James p , Stew.J.f1 o( the Funk 1. Seiler
Research Labanlory. Colondo, ancl distr ibutecl by the Quantum
Chem.ical Program Exch:ange ,
1201 11 . Romrnc:lspacher and R. Susilo, Prog. DflIg Res., 29. 41!j
(19g5).
1211 H. RommelspAcher. T. May, and R. Susilo, Planta Med.. 57.
585 (1991).
(22) I. Gynther, S. P. Lapinjoki, M. M. Air.ksinen, an~ P. Pcu...
BiocMm. Phanuocol .. 35. 2671 (1986).
123) R. D.hzly. 1. Am. Chem. Soc .. 75. 6038 (1953).
1241 A. Drossi, A. Focella. an~ S. Teilel, J. Med. Chem .. 16,418
(1973).
(2S) P. Syke.s. in A Guide 10 Mechanism in Org3n..ic Olemistry.
Fourth Edition. Longman Group u~., Lon~on, 1975 . p 25.
1261 T. R. Bosin. D. HolmSled. , A. Lun~ m .n . an~ O. Deck. in
Progr= iD Clinical an~ Biological Research. Va l 90, BelaC..bolincs
and Teluhydroisoquinolines, F. Bloom. J. Barchil.!, M. Sancller, and E.
U,wn, eds, AI.n R. US!, New YOfk, 1982. pp 15 27.
(27J P . Peura, J. Kui . and M . M . AirillcJinen. Biomed Mass
Spectram., 7. 553 (1980).
(281 O. Beck an~ K. F. Faull , Biochem. Pharma col .. 35. 2636
(19g6).
129) O. Deck. D. B. Repke , and K. F. F.ull , Biomed. Environ.
Ma.rs Spectram.. 13, 469 (1986).
130) R. Susilo and H. Rommelspacher. Naunyn-Schmiedeberg's
Arch. Pharmacol.. 337. 566 (1988).
[31) I. Gynthct, Acto CMm. Scalld.. B42, 433 (1988).
v

J. C. Collowoy'~', M. M. Airoksinen' ond J. Gynlher'
Endogenous I>Carbolines and Other Indole
Alkaloids in Mammals
Ab,lrod
Vir1uolly 011 of our perceplions enler conseiou,no.. Ihrough Ihe
chemi'lry of Iho cenlrol nervou, 'y,lom (CNSI ond Ihrough Ihi,
fen/asle medium, consdousness ,Irives lo express lself. In consid.
ring Ihe chemislry of consciousness, one moy wonder how we
facilita l. percePlion/oworeness ond why cerlOin exogenous chemi
cols hove Ihe copocity lo oll.r Ihase phenomeno. Some of rhese
psychooctive chemicols aTe opparenlly endogenic in mommols te. g.
melhylaled Iryplomines ond l3-corbolinesJ. Ihu! il seems reosonoble
lo posillhol Ihay hove sorne function in our daily lives. Ir is olso usl
as reosonoble lo presume rhal exogenously odminislered com
pounds, idenlical Of similar in kind, are octiv. vio onologous me
chanisms. From this perspedive. endogenous indoNt olkdoids ore
proposed lo calofyze periodic olteralians in cansciausness ond
Ihereby assisl in Ihe daily mainlenance af mentol healrh. Speciol
emphasis is placed on Ihe pulalive role of these endogenous com
pounds in dreoming and psychasis. and Ihe reNtvonce of Ihese
slates lo the effecls af exogenous Iryplamine derivales.
Inlroduclion
The occurrence 000 awareness of consciausneu l has b&en of inlenu'
lo out species, perhaps si nce Ihe advenl of frantol Jobos. Throughaut
lhe oge,. phonomena of Ihe miOO hove been reoognizod, explorod
ond embodied wilhin numerous personol 000 social oosm<kgies.
From relks af our prehisloric onceslan, we find omple evidence lo
,upporllhe focIlhol much of Ihi, eorly explorolion wo, ocoampli.hod
Ihrough Ihe use of p'ychooclive plon". Even oor modern sociery ocn
troce some of its neritoge bock lo andenl Greece 000 the Mysleries of
Eleusi, [WoSJOn el 01 . 1978], where o p,ychooclive ",b,lonce olher
Ihon olcohol wos certoinly in use. However, such evidence from Ihe
disJonl past le lis u.s lillle obout Ihe experience ,self ond even leu of
how il reloles lo lhe presen!.
Thi, cenlury, ond e'pec;olly during Ihe 10Sl 50 yeo", o rediscovery
of the psychedelic 2 experience b890n in Weslern cultures. More
Ihon ju,1 o hondful 01 dodicolOO explore" hove inve'ligolOO Ihe
phormolocogy of psychooclive subslonce from plonl or synlhelic
sources. N'tore recenlly. some of Ihese some subslonces hove been
idenlifled os normol componenls of humon urine 000 blood, nomely
Ihe melhyloled Iryplomine, [Wolker el 01. 1979, Roisnen 19840]
ond \<orbeline, [Airoksinen & Kori 1981, leino & Airoksinen
1985]. Allhough Ihese compound5 hove boon implicoled in Ihe
oliology of menlol illne .., Ihey oppeor in Ihe normol populolion o,
well. In oddilion, !hese somo ,ub,lonce, hovo been repeoledly
d.locled in Ihe 11.."", 000 nuid, of mony olher onirrol" p'ychOlic
or olherwise. The chemicol bosis for on obnormol sto. of mind is
commonly oeeeplod by Iho inVOCOlion 01 ondogonou, 'p'ycholo
gen,', Ollhough, li"le i, soid 01 lhe Ihoir po,sible fvnclion in lhe
normal mind. However, il js quile likely Ihal if undesirable slales of
menlol heolrh con resulr from o permonenl imbolance of endoge
nous ehemicol inleradions, lhen the healthy mind may hove o
cerlain copocity ond use for ,hese slales os welL Dreoming, an
euenlial experience. is o good example of an altered menlal slate,
one which emerge, regulorly ond ho, olllhe quolilie, 01 o holl.,.
cinorion . The origin ond purpose of dreams ore poody undersrood,
'
and wilhin Ihe process of modern culture lheir importance is eosily
overlooked. Cultures lhal inl89,ole dreoms, 000 olher olternonv.
slales of consciousness. into the fobric of Iheir everydoy lives re
eognize rhe importance of Ihis difference in perspective ond ils
polenlial lo d..pen and enhance Iheir quolity of living.
The mosl impressive ospecls of dr&oming coincide wilh ropid eye
movemenl sl..p IREMS). Since lhe mosl vivid dreoms ore echoed
by Ihi, phy'iologicol process. il j, perhap, jU'lified lo 'ugge,1 Ihol
,he visual imogery lseU moy olso bo re\ol&d lo o biochemicol
mochoni,m [Collowoy 1988J. REMS moy olsc provido !he periodic
brain slimu~lion neceuory lo insure r&COvery from sleep [Vertes
19861. Perhops some forms of psychosis resull from an overodive
or off>el drooming mechoni,m [Meurizi 1984]. Tho importonoo of
dreoming js monifesl in ils obsence; i.e. REMS deprivolion is como
pensclod by O robouOO of REMS oclivily in ,ubsequonl nighl' of
undi,lurbed .J... p in hoohhy volunl ..." [Demenl 1960]. Though il
ho, nol been demon,lroled Ihol REMS deprivolion re,ull' in idenli
fiable allernolions in woking behovior, par .se, il is well known Ihal
vivid hollucinolions ond olher behoviorol di sonden occur in humons
oher only Ihree doy, of 10101 ,loop deprivolion. Undoubledly,
however, Ihis is nollo S<?Y Ihol dreoming locb on es.senliol fundion
bul rolher Ihol drooming con oocur in non-REM ,loop. Through
dr&oming we acceu eertain aspecls of our psyche which ore nor
molly inocceuible during woking consciousness ond in Ihis sense,
dreoms shore o qualiralive eommonality wilh cerlain types of psy
chose, ond p,ychedelic ,Iole,.
Another ospecl af ehemicol eonsciausness con result when Ihe
olherwise normal person willfully salf-odminislers o psychooclive
subslonce. While !here is litrle evidence lo support o nalion Ihol
any humon society has exisled wilhoul Ihe benefil of psychooclive
substonces, il is hord lo ignore Ihe self-deslructive hobils embraced
by our own. The Iypicol diel of o modern sociery offe" muhifo
rious preparalians af e!honal, nealine, sucrosa ond caffeine Jor
od Jib consumplion. Extreme misuse of ehemicol substonces ore
exemplified by injecred combinorians of opio les, amphelomines or
cocane, Svch ponerns af ~hovior Soeem lO serve no other purpose
Ihon fulfilling sell indulgence Ihrough immediole grOlifiOOlion, ond
in Ihi. Ihe....ub.lonee. ore ex""edingly efficociou,. The u... of
poychedelic .ub.lonces i. reloliv.1y n.w lo our culture. olthough
widely uoed el...where for millennio Iheir oppropriol. place ond
fvnction in o modern SOCtety remoin uncleor. However, Iheir oppli.
eolio n o' lools for expkJring Ihe mind os well os lhe broin ore
indubilobl d.'pite polilicol r.pr.ssion ond phormocolagicol
ignoronce. The....ub"once. hove enjoyed o lang hi.lory of sociol
u....pecificolly for lheir profound effect. on Ih. p.ych. ond th.ir
nherent sofety in potterns of responsible use.
P.yched.lic sub.lonce, ond humon,
In lhe .tudy of .Ihnophormocolagy. the u.. 01 o poychedelic .ub
tonce. hold o porticular int.r." ond fosdnolion . peciolly for
tho ... locking o w.1I ."obli.hed culturol cont.xt far such expori.
.ne.,. Thi, unique doss 01 compound. l. choroct.rized by Ih.ir
obility lo reliobly induce sloles of consciousnen which are expert
cular Iev.l. wher. seemingly unreloled .trvc:tur play .imilar type,
of -music",
Frorn !h. perspectivo of argonic ch.mi.try. mo.1 chemicol. wilh CNS
octivity feoture o substiluted oromoric ring onoched fa on aliphotic
ni ~ogen. ond Ih... ore common feolur of o p.yched.lic moleeule.
Th.... feolur., ore Ihoughl lo modify rec.plar oc.1ivity in Ih. CNS by
hydrophobic inl.roc~on. ond hydrog.n bonding 01 !he receplor. from
Ih. oromoric ring ond oli pholic nitrogen oIlhe maleeule. r..pecliv.1y
(lIoyd & Andrew, 1986]. 11 i. eo'y .nough lo define Iryplomin
phenethylomines or ergol derivatives os groups unto themsetves, but
o unifying !heory lo reconcil. Ih. common psychic .flect. frorn lhe..
diverse slructural closus remoins elusive, In oddition lo the diiemma
of similar .flecls frorn dissimilar cla .... of compound, i, lhe predico
menl where chemiocb sub.tonliolly similar lo pow.rful poyched.lics
hove lilll. or no CNS ocnvity oIlheir own .
.hp~.1t(.

"I~~
0i:CCH:,
NH, } .1I.h .. " {

Ji'"'
enced onty during sJeep 01 somelimes during orar. ecslolie epipho r\lt r~I'"

oH NeH,
nywhil. owok. Simi!otilie, berw n cartoin s.Ioges of psychoses
ond p.yched.lic .tote. ore worth noling. !hough .uch onolagie. ~CH' H,CO ., om.tic:
OCH,}"n. {
hove diminished in volue over lhe posl decodes. In o brooder N oeH, N
H
MnS., out modern cullure, hove yel fa recognize how ,hese experi.
P!.iloc i n ""4'".llnc LSD
.nce. con b.nefit the Individuol ond society o. o whol . Ideolly the
(a phen4'thyl.mlnl"
poyched.lic ch.micol. hold out o hopo for understonding ond
perhops improving c.rloin o.pect. of m.nlol heolth. Asid. from
.nlight.ning Ih.ir p.ychic .Itoct. con 0110 be frighl.ning. o, Ihoy The que.lion ori ..... Ih.n. o. lo why Ih. p.ych.delics influ.nce Ih.
oft.n exoeed our frome of perlOnol ond culturol r.ferenc. How.. mind in .uch o profound woy. Th. onsw.r i. porlly in Ih. woy lhol
ver, ,he main feoluf. Iha! chorocterizes rhe psyched.lic:s is ,hei, many of Ihe Iruty psychedelic ehemica/s ore similar, and somerimes
ob;lity lo I.mpororily induc tot of mind thol oft.n Ieov. o pro id.nlicol. to ,ubSlonc produced by rh. body on o doily bosi .
found imprint on ,he humon psyche. This proves consto ni over rime Such .xogenouub.lonc ore oCQOmmoclaled by Ih. CN S ond
ond i ot Ieo.t in port. indep.nd.nt of cultur . Mor. oh.n lhon nol. lemporarity alter lhe srole of biogenic omines lo produc. Iheir
o cleor recollection of this experience remoins with lhe individuol effeels. One musl 0150 fo consider me effect ilself, since ,he mere
lang oh.r Ih. ocul. ef1ecls of Ih. drug hove po.oed. In conlro,' lo
copocity for such on .xperi.nce .uggell' lhollh. p.yched.lic .lale
mo.1 drug Ih. p.ychedelic> exhibil o high Ih.,opeulic ind.x ond
i. inher.nlly fundom.nlollo o.peel' of our p.yche Ihol o,. normolly
Ih.ir u... ho. nol been o.socioled wilh phy.lcol depend.nc.
inoccessibl. during Ihe woking pho .. of our liv. Thu und.r
A ropid ond I.mporory induction of lolerone. (which i. nol compe'"
oppropriote circumslonces wc:h ehem icols moy 'empororily ollow
soled by incr.o.ing Ihe do...) i. 0110 chorocle,i.lic in Ihi, doss 01
on individuol de.p ond brood ooc... la Ih. mind.
drug' [Joff. 1990]. Pouible exceplion. ore of N.N-dim.

Ihyllryplamin. (OMTJ ond p.rhops s.m.lhoxy-dim.lhyhryplomin. (.5

MeOOMTJ. wher. r.peoled use doe. nol reodily indu"" 100.ron"".
Namenclatur.
Thi. moy be due lo Ih.ir remarkobly fo.1 meloboli,m. Bolh of lhe...

hav. been id.nlified o. normal comp.n.nls of humon blaod. ond

Since chemi.lry wiU ploy o cenlrol rol. in Ihi. orlicle 11 wiU be
Ih.ir .ffecl. will be diseu.oed lol.r.
necessory lo brieRy review the nomencloture pertinen! lo Ihe moleeu
Th. mechonl.m of oclion for poyched.lic ch.micob ore .,iU undeor.
les in queslion. The main chemicol f&atures which tryptomines and
Currenllheori discu.. the ...fleets o' rulling from complex
~orbolines Ipesl soore in common ore o heterocyc:lic oromatic
interoctions w ith vorious receplor subtypes of Ihe serotonergic ring (indol.) wilh on olipholic nilrog.n (.Ihylamine) onoched ollhe
'Y'I.m (GI.nnon 1990. McK.nno .1 al. 1990. Apud 1991. Oeligo. Ihree position of !he ringo Th. indole ring is o compone nI of mony
ni '0/. 1991. Nichol. el 0/. 1991. Slrossmon 1992]. wher. o biooc:tive subsfanc:es in noture . Irldole is chemically oromolic ond
certoin populolion of receplor .ubtype. may bIimulaled while comprised of O hexogonol (phenyl) ,Ing fuoed lo a penlogonol
onother ore inhibited , These .ffecls ore nol unlke o note, or rolher pyrrol. Th. nom.nclolur. for indol ond tryplomln i, foirly
chord. of mu.ic offecling Ih. ph.nom.non of heoring in Ih. mind. direct, where numberjng begins 01 Ihe oromalic nitrogen otom on
likewi .... o chemicol moy play Ihe... nol or chord. Ihrough o Ihe indole ond proceeds caunterclockwi.s.e oround ,he ring 01
voriety of receplor .ubtype. lo off.cllh. ph.nomenon of conseiou .. .hown. (Th. inlerfoce b.rw.en Ih. ph.nyl ond pyrrol. ringo i. nol
ne... How.ver. Ihi. onology i. e.peeiolly bewildering on Ih. mofe. numbered, sinee il con not OGCommodole rurther substilution).

2
 The word , hormol' is opporenlly of Arobic origin ond hos been
used lo refer lo Ihe plonl, ond espoeioUy ..eds, of P8gonum
hormo/o since oneienl times . The oldesl known deseription of Ihe
plont wos probobly token from eorHer Arobie lileroture by Diosco
rides in his compilolion of De mOlerio medico, o firsl eenlury texl
on medicinal herbs [Gunlhor 19341.
Indol~ 5~Methoxydlm~thyltryptamme The hormolo olkoloids, excepl for horman, were firs! discovered in
Ihe seods of Pogonum hormo/o [Gbol, 18381. HOi'moline ond
Tryplomines ore ndole slruclures hovng on elhylomino ottoched lo
hormine were 01$0 idenlified 10ler os componenls of Bonis/eria
polilion three on Ihe ring o Subslituenls on tha elhylomine nirr0gen
(sic) [Hachsroin & Porodios, 1957]. Th. hormolo nomoncloluro
are designole'd by Ihe nome of the substiluenl, preceded by ' N ',
endures os o rele 01 aorly explorolion, Ihough rheses eommon
This is illuslrOled for 5-melhoxy-N,N-dimolhyltryplomine 15
nomes ,eveo! litlle of Ihelr moleculor choraelerislies. AI$O, many of
MoODMn, which is o Iryplomino wirh Iwo I'di') melhyl
rhe~ subslonees shore muhlple idenliries. Fot axample, hormoline

(-CH,) groups bolh on Ihe .ame olipholic nilrogen (N, N) ond o

is obo known by Ihe eommon nomes 01 hormodine, hormolol
melhoxy .ocH 3) group 01 posilon five on Ihe ring o Two olher
methyl ether or O-melhylhormolol. Hormine is oquolly nolorious
imporlon! me1hyioled trypromines discussad in ,his onide ore
wilh nomes sueh os baniste,ne, yogeine. teleporhine 01 leueohor
N,N-dimelhyltryplomine (DMn ond 5.hydroxy-N,N-dimelhy~
mine. Apporenlly these subslances meonl diHeren! Ihings to the
Iryplomine (5HODMT or bufolenine).
mony diHeren! people involved in Iheir modern tooiscovery. Also,
The nomencloluro of Ihe ~Cs (belcxorbolines, 01>0 known o. 2
011 of rhe hormolo olkoloids ore a merhyl subsrilu!ed, rhol is they
eorbolines) is ohen len diraer, ond olmosl o discipline unto itself. To
hove o melhyl graup al posilion one. lorer, ~Cs wefe diseave(.d
begin, Ihe PCs con be 80sily divided inro IhrHl moin slructutol
which loeked rhis subslilurion ond il beco me necessory lo nome
calogorio" lhe fvUy oromolic jkorbolinos (JlC.1. 3,4-dihydro-'
them os norhormolos. 'nor' meoning 'wilhoul O rodieol' [e.g.,
methyll. In Ihis way, nornarman ~ ~<arboine. Mony olher gymno
corbolinos IDH~Cs) ond Iho fuUy >olurored 1,2, 3, 4-!0lrohydro-j}

corbolino. (TH~Cs) . Tho chemicol slrucluros for ~<orbolino (llCl ,

sl ie leols of nomencloture were implemented over Ihe yeors lo
dihydro-jkorboline ond 10lrohydro-~rboline (TH~Cl oro os
preserve Ihe hormolo nomeso~e , ond mony J}Cs con be nomad
follows ond the numbering system for 011 ~Cs remains eonSk)nl.
wilhin Inis syslem . However, Ihe hormolo nomenclatura is rorher
eumbersome for PCs wnien o'e nol o.-SUbslituted, ond in proctica

,Q-CN
6 '5 " 3
onlyo few rKs cont inue lo be !eferrad lo by ,ha;r hormolo nomes.
Tne mosl specific, ond leosl obviaus, wcy lo nomo cny corbalne
N (a. ~, r or 5) would bo lo use Ihe IUPAC (Inrernorionol Union 01
H
Pure and Applied Cnemlslryl syslem of nomencloture, which coll,
DH~C THBC Ihem 011 pyridoindolas. Tnus Ihe common no me of hormoline
Iron,loros lo 4,9-dihydro-7.morhoxy-l.merhyl.3H.pyrido[3,4
The mosl woU known ~s oro Ihe hormolo olkoloid., i.e. hormolino, ~lindole. From ,his example il is eosy lo ~e why 'hormoline' hos
harmalol, hormine ond hormon , These ore shown lo iUuslrole Ihe endured os Ihe name 01 choice . However, some pyridoindoles con
slruclurol similorities between Ihe originol hormolo olkoloids, ond lo bo clossiliod os ~<orbolino, (~C,J. where rhe PCs ore Ihe fvlly
provide Ihe;r nomes in lerms of Ihe J}C clossificolion s)'3tem . Como oromOI;c ,lrueruros 'oUowod by dihydro-jkorbolines (DH~C,) ond
mon nomes for Ihase compounds should be usad whenever po~i. linoUy rho fuUy >olurorod 10lrohydro-jkorboli nos (TH~Csl. In Ihi,
ble, sineo Ihey ore mosl fomilior lO lhe lorgesl number of peoplo, system, hormo!ine beeomes 01 leo sI monogeoble os 7.melhoxyl
Ihough 01 100S! pouing reference lo Ihe ~ nome should bo mode merhy l-3,4DH~C, or 7,MeOI MoDH~C, ond hormine becomos
lo nsure occurocy. 7.melhoxylmerhyl~C , or 7.MeO-IMo~C. This is cerloinly le"
complicoled Ihon rhe IUPAC sy"em, rhough odmilledly ,tiU more

H'COQ-ON HOQ-O.

eomplieoled rhan simply using Ihe Common no mes.

TH~Cs hovo olso beon collod Iryplolinos [Borchos 0101. 1974,
Wyoll el 01. 1975, Kollor o, 01. 1976, Rommel'pocher el 01.
~ eH, ~ CH,
1976, Elliol & Holmon 1977, Youdim & Opponheim 1981, Bed
Hbrmallnl Harmalot
& Foull 1986 ond Pouro el 01. 19891 , since mosl of Ihe ~Cs found
(7-methoxy-l-methyl-OHt.C) (7-hydroxylm.h t I Y'DH~C)
endogonously ond in Iho diel ore TH~Cs [Mcl.aoc 1961, Borchos
810/. 1974, Hsu & Mondell 1975, Peuro 810/.1980, Airoksinon
& Kori 19B lo, Kori 8101. 1983, Bed & Foull 1986, Bosin 0101.
1988, Bosin & FouUI989, Adochi 010/. 1991j . Tho moin odvon
loge lo Ihis syslem of nomencloture is Ihotlhe noming ond numbe
ring resembles Ihol of Ihe Iryptomines from whieh they were foro
HD(min~ Hilrmon medo However rhis lorminology is limited lO Iho TH~Cs ond is nol
(7-methoxy-t-mcthyl-lC) (-m<thyl-OC) roodily opplicoble ro DH~C, or rhe fully oromolic ~Cs. Tho Iryploli.

3
ne syslem of nomendolure hos mel wilh somelhing less Ihon
universol oeceptonce ond its usoge seems lo hove diminished
sinco ils inceplion. On roro otalsions, TH~C ond l-Me-THPC havo
been tefered lo os noreleogine ond eleogine, respectively.
Anolher trend in noming new compounds al o series ulilizes on
olpho-numeric syslom (e.g . lSD-25) wilh o logic ohen opprecioled
by o limited number 01 people 01 ony given lim. Though mor.
versolile rhan whimsicol eommon no mes, ond certoinly more
eompoe' Ihon rhe IUPAC syslem, Ihis syslem remojns mosl usefullo
lhe synlhetic orgonie ehemisl ond is oceosionolly encounlered in
r.lerence lO synlhelic products.
In mosl cosos il is probobly besllo r.fer lo Ih. mojar PCs. ond
olhe, ehemicols, by Iheir mosl populor nome while olso noming
Ihem wilhin o single sysl.m copobio 01 importing o specilic ond
informolive nome. In this woy confusion moy be ovoided while
inlerdisciplinory eommunieolion is foeilitoled.
Genesis of lhe rronsmelhylolion hypolhesis
To underslond Ihe normol role of endogenous psyehedelies il is
impor1onllO know somelhing of Iheir discovery. Since Ihe eorly
1950's. r.soorch.rs in ,he lields oF mon'ol hoolth hod olreody
proposed Ihecries concerning ,he ehemieol nolure of mentol
disorders, porticulorly schizoph'enio. One prominenl omong Ihe$8
has been ,he 'dopomine hypo'hesis', which .ugges's ,ho' hyperoc
livity in dopomine'gic syslems resulr in signs ond symploms of Ihe
diseoso WyO" 1985). Thls typo 01 psycho.il is choroc'erized by
symploms of poronoio, ond resembles omphelomine induced
psychosis fa sorne degree. An inleresfing slruelurol similority exists
belween omphelomines ond eolecholomines sueh os dopo mine,
,hough unlike poronoid schizoph,.nics. dodico'ed ompho'omine
users typicolly reloin some insight ond ore usuolly owore ,hol Ihei,
Feolings or. drug induced.
ships eonloined lohoeeo for rhe future of Europe ond Waslern
Ano,her ,h.ory wos dovoloped o. Ih. 'Ironsm.,hylo'ion hypolhe
civilozotion, o highly oddiclive subslonce virluolly unknown for i1s
sis', which focused primorily on Ih. origin. 01 hollucinolory psy
visionory quo/iles.
chasis [O.mond & Smylhi.s 1952). Bosed on Iheir p.ycholic~ike
In ,h. mid50s, Iho Firsl ovidonce wo, publishod lar Iho lormolion
experiences wirh mescoline ond othet phenethylomines, ,e$8or
chers propased Ihol sueh o 'schizoroxin' or 'Ioxic psychotogen'
could r.suh Irom Ih. endogonous m.lhylolion 01 cerloin biogenic
omines. Notng the structurol similorilies between mescoline ond
some of the neurolronsmitters, ecrly reseorehe,s seorched for
melhyloled d.rivo'ivos 01 dopomin. ond noropineph!ine whlch
could occounl for Ihe hollucinolions ond defusons symptomotic of
s.chizophrenio. This work eventuolly resulted in Ihe idenlificotion of
3,4-dim.'hoxyphenelhylomine (DMPEAI. or Ih. lomou. 'pink .pol'
In Ihe urino 01 polienlufferng from schizophronio [FriodholF & Ihol enzyme. in robbillung could produoo bufalonine, DMT ond
Van Winklo 1962). However, DMPEA wo. 10le, lound in Iho u,in.
of normal .ubjecls 100. ond somelimos n.vor lound 01011 (Kuehl el
01.1966. Bouhon 1971). Anolher d,owbock far Ihis version 01 Ih.
Iron.molhylolion hypolhosis wos Ihol DMPEA is nol psychoocliv.,
ovon 01 hlgh doses [Shulgin & Shulgin 1991] conlrory lO earlior
roporls [Smylhios 1960). Some yeors lolor Ihis somo lino 01 Ihin
king wo. opplied lo Ih. in vilro biosynlhosis 01 moscolino by livor
enzym.s [Friodhoff el 01. 1972, Rosengorten & Friedhoff 1976).
4
However, mes.coline has yel to be determlned 01 on endogenous
component of ony onimol.
Exomples such os Ihese ore o reoeeurring feofur. of pioneering
rosoorch. where crucial deloils 01 on onolyticol m.lhod moy be
inodvertonlly leh oul 01 lhe publi.hod procedure or Iho rosults were
ocluolly orlllocluol ond unr.cognizod lO be so ollhe lime. Howe
ver, sueh onomolies ore Ihe' unfortunole byprodUCls oE initiol
investigotion rOlher thon peor seienee. Old geogrophicol mops
oplly ond eonsislently illustrole this point. Sueh O Iroil moy be O
common feolure to 011 forms of explorotion.
Oeeurence al melhyloted tryplomines
Publishod ,eporl. on ,ho p'ychoOClivity 01 melhylolod Iryplomino.
bogan lo oppeor in ,h. mid 1950'. (Fobing & Howkins 1956.
Boszormnyi & Szoro 1958]. saon oher their rediscovery in
psychooc'ive snuffs 01 Ihe New World li ... Virola ond Cohobo
snuH'I. Bulo'onin . 5-MoODMT ond DMT or. me,hylo'od 'ryplomi.
nes which hove boon faund in Iho _d, 01 Anodononlhoro p<lre
grino IFormorly Pipladeniol, o mojor compon.nl 01 Cohobo snulls
[S'romberg 1954. Fish el o/. 1955. Holm,ledl ond lindgreen
19791. whilo Virola snuffs havo lockod dolecloblo omounls 01
bulo'onino [Tarros 01. o/. 1991)
Bolh Cohobo ond lobocco .nuHs wore once used Ihroughoullh.
Corbbeon, ond Iheir use wos firsl described lo rhe Europeons by
Frior Romon Pone in 1496. 1I s templing lo muse on rhe euriosity
oF on oxpoclonl monorchy. piqued by lhe discovery 01 o substonce
copoble of inducing visionory experiences, since the power lo
heol ond prophesize would bestow on oddi'ionol odvontoge to o
pe,son in power. In subsequent yeors, how8ver, the Europeons
Froquonlly conlused Iho oboriginol use 01 Cohobo wilh Ihoir use 01
lobocco snuffs. C,cumslonliolly, perhops, the co'go oE r.turning
of burotenine, ond its monomethyl dervotive, from Ihe neulronsmi"
lor 5HT [Bumpus & Pog. 1955). Whilo Ihor. l. slill some doubl o,
lo wholh.r bufolenino is Iruly psychodelic (Shulgin 1981), mosl
studies show bufotenine lO hove o 51ron9 ond ropid onsel of
psychooetive eHeets, when odmisle,ed intravenously, which o'e
similar in durolion lo DMT ond 5MoODMT (Turner & Morli.
1959. Bickololo/. 1976, Mcleod & Silorom 1985).
Inle,esl in the identificoton ond quontificolion of endogenous
Iryp'omlnos incroo,ed in Ih. oorly 1960'. oher Ax.lrod observed
Ihei, monomelhyl derivotives from primory Iryplomines by using S
odonosyl molhionin. (SAMios o molhyl donar [Axolrod 1961 ond
1962). lolor o similar .nzymo wo' discovored in lhe CNS, Ihe
highesl concentraran! occuring in ,he broin slem ond lhe leosl in
lhe conicol oroos [Margan & Mondoll 1969 ond 1971). Thoso
linding' lurlhorod Ihe hYPolhesi' 01 o role lor m.lhyloled Iryplomi
n.s in Iho origins 01 psychosis [Gillin 0101. 1976).
Also in lhe eorly 1960's, Pollin ond ossociolo. discovered lhol
symploms 01 schizophrenio could be e.ocarbaled by odminisl.
rlng iproniozid (an anlidepressont ond monoomine oJ<idose
(MAO inhibilor' wilh melhionine, on essenliol omino ocid ond
melhyl donar [Pollin el 01. 1961, Alun el 01. 1971 l. This work wos
loler conlirmed ond reviewed [Airoksinen & Mclsooc 1967,
Cohen el 01. 19741. In olher sludies, il wos found Ihol rese'pine,
in combinaron wilh MAO inhibilon, iniliolly Increosed endage
nous Iryplomine concentrorions os well os psycholic behovior
(Brune & Himwich 1962, Berlel el 01. 1964, P.cheidl el 01. 19661.
This, plus Ihe locl Ihol severol 01 Ihe melhyloled Iryplomine. we,e
known lO be psychoactiv8, led mony investigorars lO explore rhe
putativO' connection between psychO$i$ ond endogenous Iryplomi.
nes well inlo Ihe 1980's, while Ihe nolion 01 endosenous ring
subsiluled phenelhylomines become o moner of history.
Th. bio-tronsformotion of Iryplomines lo melhyloled Iryplomines is
Ihoughl lo proceed os described below.
o R.. H R t:H,
~OH ~NH ~NCH'
N --
N -
N
H H H & Van Vunoki. 1978, Risanen & Krkkinen 1979, Si'orom el
Tryplophan R H,CO. S M.O! R H,eO. ~ ' M,OCMT
HO. S"OIOO", HO, 8ulcunlOf
H, TryptamlO' H. DMT
Tryplophon from Ihe diel i. enzymolicolly decorbo.yloled lo
Iryplomine (R.HI. which con be melhylo.ed by N-me.hyl. ronslelo
ses (NMn in Ihe presence 01 SAM lO lorm DMT. Tryp'omine ond
DMT ore nol known lO be hydroxylOled in momma[ian liS3ues lo
se,olonin (5hydro.ytryptomine, 5Hn al bulo.enine (5-hydro.y
N.N-dime.hylrryplomine, 5.HODMn, re.pec'ively. Ins'ead, Ihey
ore lormed ohe, Ihe hydlo.ylo'ion 01 .ryp.ophon '0 5-hydro.y.ryp
.ophon ond ils decorbo.ylo.;on lO 5-HT, which is N-melhyloled '0
bufotenine. Thraugh Ihe the enzymalic aelians aE 5-hydraxyindole
Om hyl-.,onsfe,ose (HIOMn, sero.on;n ond bufotenine moy be
lur.her methyloled lo 5-MeOT ond 5-MeODMT, r.speclively.
The odminist,olion 01 DMT is known lo induce o florid display 01
visual firewarlu, lasting anly o few minutes, and repetitiva use has
no' been .hown to develop loleronce lo Ih;s effec' [Szoro 1961,
Shulgin 19761. The phormocokinelic effect. 01 5MeODMT (o
more potenl .ubonce ore similar to .hose of DMT, ol.hough
locking in visual phenomeno . In"eod, Ihe effects 01 5-MeODMT
ore considered fo be primarily emolive imagery, ralher Ihan
visual, with reporfs oE mentol SIOleJ resembling neor death exper;
encas [Ge ..ner & Poge 19621. Introvenou. odminislrolion 01
buFolenine shows o similar phormocokinelic profil., wilh some
indicolions 01 psychooctivily, bul signi~conlly locking in Ihe ellects
so chorocterislic o, the other two chemicals when given under
similar circumslances. Alllhtee af !hese tryptamines afe eHectively
metabalized by several roules ond ore quicldy eliminaled Iram rhe
plasmo. While .he psychoocliviry 01 DMT ond 5-MeODMT ore nol
in dispule ond Ihey cerroinly penelrole Ihe blood broin borrier
quile easily, Ihe hydroxy SUbsliruent an bufarenine prabably dimi
nishes its ability lo enter rhe broin when odminislered exogenous-
Iy. However, bufotenine moy hove oclivity jf il is formed neor ils
si te of oclion in the broin. II js opparenlly o polenl psychoaclive
JubJlonce when enough reoches Ihe CNS oftef inlravenous admi
niJtrolion, though Juch investigol,ons Ofe limited since hgh doses
of bufolenine ore ohen complicoted by onoxemio and alher
serolonergic eHecl. (Fobing & Howkin. 19561 . Perhop. ony
psychooctive eHects nom exogenously odmislered bufotenine
resuh from ils ;n vivo convenion lo 5-MeODMT vio Ihe en~me
HIOMT.
Over Ihe ensui ng yeors, rhe delecllon 01 bufotenir-.e ond olher
melhyloled Iryplomines in human fluids wos confirmed ond refuted
under O voriety oE circumslonces (Norosimhochori el 01. 1974,
Guchhoil 1976, Chri,'ion .'0/.1977, Corbel! el 01. 1978, Wo~
ker el 01. 1979, Borkel 1981 l. Ir seems .ho' mony loc'ors con
influence Ihe onolysis [H,yhorczuk el 01. 198610. well o. on
individuo!'. somple [Pscheid. el 01. 1966, Oon elol. 19771.
Unlike Ihe olher methyloted Iryplomines, buforenine is primorily
excreted in the ur;ne ond has been found in samples fram healrhy
volunteers ond in unselected psychiatric palienls who ware under
going Irealmen' wilh vOlious drugs [Krkkinen el 01. 19881.
These results ore in ogreement wilh eorlier sludies which hove
u.ed go. chlomo'og,ophy-mo ...pec'romelry (GC MSJ [Riceberg
01. 1983, Risanen 19840&bl . The difference. found by K,kki
nen el 01. beIWeen Ihe heollhy valunleers and psychiolric pOlianls
were nol os graol os individuol dtfferences wirhin Iha groups, ond
.he octuol omounls of bufolenine moy hove depended more on
diel, gut bocteria ond medicolion Ihon individuollemperomenl.
The5e reparts roulinely oss.ociole endogenauJ bufolenine wilh o
variety al human charocleristjcs such os occoJionol desfructive
ond violenl behaviar, anxiety. palanoio, misperceplions ond
5uicide. Whal Ihis soys fo, nafmol humans is une/eor. A ponern of
oscribing foulls lO an endogenaus psychooctive 5ubsronce, while
Ignoring ils putative implicalians in normal behoviar, is typicol of
Ihis ero.
From .he eorly 60's un.i! .he eorly 80' . o ha 01 publicolions
oppeo,ed on Ihe biogenesis 01 me.hylo.ed tryplomine. which
eilher canfirmed, denied 01 ,econfirmed Ihese discaveries while
ignoring o possible role in normal behaviar. A rore exceplion wos
o publicolion by Jocobs ond Trulson in 1979, who hod consultad
Ihe eorly lileroture 01 Piolo ond Ari,'o.le lO link hollucinolory
phenomenon lo dreoms, p5ychedelics and psychosis. Apporenlly
Ihe loner poir or reseorchers cauld nal agree on 'whelher dreoms
and Ihe hollucinotions associoted wilh diseose orase from Ihe liver
al .he heor!' Ihough 'Ihey were in occord wi.h Ihe beliel Ihol bo.h
were producls of the some process'. Jocobs & Trulson exlended
this line of reosoning by including drug.ndvcad hollucinotions
along wi,h dreoms ond cerlain mentol diseoses, ond suggesled
Iheir effecJs lO be direclly attribulable ta neurochemicol mecho
ni5ms common lO olllhree. This speculolion resulted from thair own
re5eorch on ,he CNS in general, ond Ihe serolanergic system in
particulor. An excellent review af ,he literoture concerning other
a5pecls al the similarities between Ihele th,ee sloles of mind wos
publi.hed by Fischmon in 1983.
inl"9,olon, CMlrflol ku ",!\d.t!'O";tog pkJnu ol'ld c",lNr. N. 5/1995 5

Occuronce 01 karbolinoo nin. liko mony o.h., ~C., pinolino inhibilo MAQ.A Udonlri.nd
1958, Ho 1972. Buckhollz & Boggon 1977, Mellor & Guho
In.oro,' in .he karbolino, (JlC') gr.w Irom oorlior work wi.h .ho 1979, Fullor 01 0/. 1986J ond ho. Iho oddilionol prope'ty 01
hormolo olkoloid" ond lo'er wilh o.hor d.rivo'ive,. In .horly Increosing broin concenlrolions of 5HT by inhibiling ils (reluptoke
1950's il wos olreody known Ihal severo! omines, omino ocids inlo prooynoplic nouronol'.rminol. [Ho 0101. 1972b&c, McI$Ooc
ond pep.id., in .ho body could provido ,ub,'roteo lor .h. non.nzy 1972, K.llor 010/.1976. Rommol'pochor 0101. 1976. Elliol &
motic lormolion 01 odducls wilh oldehydo. under phy,iologicol Holmon 1977. Mey.' 010/. 1978, Airoksin.n 010/. 19780&b.
cendilion. (Wholey & Govindochori 1951] o.g ., Ih. Iormolion 01 1980, Tomo. 010/. 1979, Buckhollz 1980, Youdim & Opp.nhoim
~C. Irom Iryplomin . A, proviou.ly men'ioned, ~C. oon be 1981]. It ho, olso be.n .hown lo b.hovo o. O hormono [Airo"'i.
lormod ondog.nou.ly Irom primory Iryplomino uch o. Iryplomi nen el 01. 1984] and its successfullriliotion has rev.oled sp6cific
no, IOrolonin (5Hn ond 5-molhoxytryplomino 15M.OTI [Airoksi. binding sires far pinoHn. in ,he pineol, od(.no~ ond vorious
nen & Kari 19810). Their identificarian os nOlurol componen!! of r.gion. 01 Ih. broin [Airo inen 010/. 1993. Collowoy 010/.
mommolion li!sves ond fluid, fUllhered Iheir inveslgolion os 19931. Olhor .ndogenou. ~C. hovo baon id.nlif.d. Ihough Ih.i,
biologicolly importon! products. Th. mosl inleresling feo tu res ol funclions hove yel lo be 500 Ihoroughly inv.stigol.d os pinoline's.
rhes-e compounds ore found in their jncr~ibly broad speclrum ol A .ch.mo lor Iho probobl. biooyn.hio 01 pinolino ond 6-HQ.
biooClivity [Airobin.n & Kori 1981b] ond in Iheir po.onliol for TH~C i hown below.
5)'nlhelic voriety.
From!ho oorly 1960'. un.il tho 10101970'0 o mojor c,ilici.m io.he HO HO

onoly.i. 01 ondogonou. ~Co cool.red oround Ihoir ortiloctuol lo,mo Qi'NHl ~ ~NH
1100 Irom oodogonouo Iryplomln.. du,ing !ho somple pr.pero~oo. ~ ~ ~10MT
Thi. possiblity woo ....ollolly olimino.ed by Iho oddilion 01 sem~ 5-HT "" '-HO-THItC
HJCO~
cerbezid. lo .ho sompl , lo trop ovoiloble oldehydo. ood by .he
inlroduction of deuleroled Iryplomines os substraros fe, the~ polen
I NA T HIO~ ' - ( J........ NH
liol reacrons. If arlifoas ware formed. !hen deuleroled ~s would
/ "H
1HIOMT cycln..
olso bo delecled by GC MS [Bock & Foull 1986J.
H,CO~ H,CO~
"--<.) H"'rO ____ "--<. N
In o,d., lo, lhe oliphollc ,idechoin lo cyclize. lhe Iryp'omloe '&qui. ~ NH I
r., on oddtionol corbon otom to COmpMtle lhe dosule and form o
H eH, H
[Xorbelin. Thi. oddilioool carbeo i. ovoiloblo io lhe Iorm of Mfl .. tonln t.-Meal
oodogonou. oldehyde. produced In silv ji .. Iormoldehyd. or
ooololdehydo) (Wo.lo,n & Ozburn 19491 o, Qke.o ocid. (glyoxylic
or pyruvic ocid) [Gynlher 010/. 1986. Suoilo & Rommol.pochor Melalcnin derive! from 5 HT Ihrough rhe enzymolic aClions of
1988]. This reodion is aften JO favorable under ocidic ccodilions serolonin N-ocotyltron,lerooo (NAT) ond 5hydroxindol...o.melhyl
Ihol much 01 Ihe oo,lier wo,k (1960'.. lo.e 1970'.) dedicoled '0 .ho tron.lerolO (HIOMn. while 5MoOT ro.ullo Irom .ho onzymo'ic
idenlifico.ion ond quontilolion 01 mOlhyloled Iryp'omino, hod pro hydroxylolion 01 5HT by HIOMT. Tho cyclizolion 01 5HT lo 6-HQ.
bobly delecled THJlC' which were un resolved from Ihe Iryplomine. TH~C, o. well o. 5MeOT lo pinoline, probobly occurs Ihrough O
in Ih. onolyticol scmpleo. oinco lhe onzyme. which Iorm me!hylo.ed non~nzymolic condensolion 01 rhe Iryplomines wilh glyoxylic
Iryplomine, in vilro con olsc lorm THJlC.lrom Ihe some Iryplomino. ocid. lollowed by .heir decorboxylolion '0 Ihe corro.ponding
Crook, el 01. 1986]. These ond o.her lechniool d'flicultie. olsc THpC o, neulrol pH . The biooyn.h io 01 THpC would be .xpeclod
bogon lo dimini,h in Iholole 1970'0 wilh Ihe opplicolion 01 beHor lo follow o similar roule in Ihis ,che me, by replocng 5-HT wilh
ch,omologrophlc sepo,olion .ochniqu.., deule,oled stondord. ond Iryp'omino ond 6-HQ.THpC wi.h THpC.
!he odvent of se~live Ion mo" speclrometric delIKlion. Th. natural oecurrence af pes has widened Ihe opened door to
.,Adrenoglomorulokopin" (6-methoxy.l.",elhyl-TH~C) wo. Ihe f,<s1 i> speculotion on Ihe puloliva roles of indole deJivorive, in physiok>
oorbolino lo be idenlified in mommolion ~ ..ue [Forrol & Mclsooc gicol ond p.ychologicol mochoni.m . Although Ih. relol.d hormo
1961]. lheugh Ihi. finding he, nevo' been ve,ified. In .hel .ome yeo, lo alkoloids were originolly described os vision inducing [Ajando'
il wos shown lo be formed in vilro under physiologicol condi'ions 1762J, .hey opporently 10cK Iho p.ychooclivity so choroclori.tic 01
[Mc1sooc 1961] . l. i, quile likely lhel Ihe ocmpound idonHfied o. o Ihe psychedelic Iryplomines. Hormalin. in particular has often
normal componen! of mommolion ti$Sue WQS octvolly on ortifoct in been de.cribed o. 'hollucinogenic' [Noronjo 1979]. Ihough

Ihe $Omple p,epo,olion. Le.. Ihon two decode. Ioler o .imilo, como corelul .Iudy ho. ,hown Ihi, compound lO b. locking in p.ychode

pound, plnoline (6-molhexy-THJlC), wo,Ioblished lo be o noturol lic effocl, [Shulgin 1977, Collowoy 1993] ond lolor reviow,loil

ocmpenenl of Iho humon pinoolglond by GC MS, ond Ioler delerm~ lo menlian such offecl, far hormoline ar ony of ils relolives [Holm

ned lo exist 01 conoenkolions .imilor lo lhese of melolonin Kon .'0/. .Iedl 1982J.

1983J. Thi, somo report .ugge.led o ciroodion ,hythm Ior pinolino in Ropor" indicole .ho. 2.me.hyl~C. moy ploy o rolo o, endogonou,

lhe pinoolo of chicken. which wo. in pholO wilh !hol of mololonin foclors in Po,kin",n', diseoso [Malsubo,o 0101. 19920&b], ond il

(i.o. incroosed levol. 01 nighl ond decreo.ed level. during Ihe doy). is worth nating rha! hormine WQS once on importonl medicine used
Pinoline, like melolonin, i. probobly o melobolic producl 01 ,erolo lo Ireol.hi. di.ooseSonchozRomoo 1991]. O.h.r loxic ~C. moy

6
be formed during Ihe preporolion of certoin foad. , Wokoboy.hi .1
01. 1983), olsa worlh no/ing ore Ihe severol 3-olkyl-~C. esler (e.g.
3-nbuty~~C<orboxylole) which ore inverse ogoni'l' 01 benzodio
zepine binding sBes where 'hay oc, os onxiogenic ond procon
vulsonl ogenl. [Broe.lrup el 0/. 1980, Peo el 01. 1986).
Anolher oreo 01 inve.ligolion for Ihe IlC. ha. locu..d on Iheir
formarion from raaelions between tryptomines ond ocetoldehyde,
lhe primory metabolit. of ethonol consumplioo. Th. ove roge
olcoholic conloins ond .xcess ocetoldehyde, wh.r. 01 leosl o
porlion would be ovoiloble lO reocl wilh Iryplomine. lo form 1
melhyl-TH~C. (I-Me-THIlC,) , Since lhe phormocologicol profile. 01
l-Me-TH-IlC, differ Irom THIlC. [Rommel.pocher el 01. 1992),
sorne reseorch has be.n direeted towords ,he investigolion af rhis
pOlonliol link in olcoholi.m [Myors & Molochior 1977, Peuro 010/.
1980, Tuomi.lo el 01. 1982, Rommel.pocher el 01. 1984, MoI.ub
oro el 0/. 1986). Ahhough olhonol diluled only wilh wol.r wo.
u.ed in Ihese .Iudi , horman (1-Me-IlCJ ha. bo.n lound lO bo o
nolurol compon.nl 01 olcoholic boveroge. [80.in el 01 1988). ond
1Me-TH~C ha. b...n idenli~ed o. o nOlurolly occurring Iroce
omine in rol broin ond lung [Bosin & Foull 1989). Th. reoClion
sch.me below .how. Ihe condensalion of glyoxylic ocid ond
oce'oldehyde with tryptomines andolIustroles ,he slrveturol diff.
renco. bolween Iho corre.ponding THIlC. ond l-Me-THIlC .
respeclively. Pyruvic ocid moy 0150 reocl in vivo with Iryplomines
lo form I-Mo1lC, [Rommel.pocher el 01. 1984, Gynlher el 01.
1986, Su.ilo & Rommel.pocher 1988), Ihough Ihi. reoclion i.
10th.r slow in vitro, under physiologicol conditions [Colloway el
01. 1994) ond oceloldehyde con be obloined from pyruvic ocid in
vivo .
Q-CNH
N suggestad Ihalo mechonism nol unlike Iho! oF Ayohuosco's occurs
H
R Hleo, P,nolln~
HO . tI-HO-THBC
H. THf\(
R H,CO; 5-M~OT
HO. Scrotonln
o
H"CH,--
RQ-CNH N
oClivity of molhyloled Iryplomine. by blocking Iheir maloboli.m.
a"tald~hyd~
K; Tr y ptamln, H
R H,CO. I-M~'pinolln,
HO, tI-HO-l-M~ - TlleC
H; I-M~-TH~C
~rbolines combined wilh Iryplomines
It i. well known Ihol ~C. polenliole Ihe oclivity of mOlhyloled
Iryplomines, ond rhis is cerloinly rhe mechonism of oclion behind
Ihe profound effecl. of bovoroge Ayohuosco [McKenno 810/.
1984, McKenno & Towers 1984, luna & Amoringo 1991).
Ayohuosco hos been used in Amozonion cultures since prehisloric
rimes lo ollow individuals occess lo tronscendentol experiences.
The p-corbolines in Ayohuosco ore obtoined from species of
Bonis/6riopsis while DMT is primorily obloined from Psycholr;o
viridis ond togelher Ihey ocl in o unique synergy to form on orolly
active beveroge . The$8' visionory experiences lend lo imporl o
holistic vision lo Ihe individual ond often o recognilion of rhe 'true
self'. A profound r.cognilion of deolh o. on importonl port of life
is nol uncommon ond is offen experienced os o deoth/rebirthing
of rhe individuol. These powerful impres.sions uhimolely extend inlo
rhe community fobric lo engender o cohesive socioreligious nel
work [Andrilzky 1989).
Recen' investigorons inlo Ihe identity of Somo, os depicled in !he
oncenl V.dic lilerolure ond poroll.led in Zorooslrionism os Hao
mo, hove implicoled hormolo olkoloid. from P. horma/o (ond
hormoline in porticulor) o, Iho oclive componen" [Flonery &
Schwortz 1989J . However, il seem. unlikely Ihollhe efleel. of
Ihew olkoloids con occount for Ihe fontastic repurolon of Ihis
mythic baveroge. Unforlunolely rhe oulhors rely on dubious phor
mocologicol dolo lo support hei, condusion of hormoline os the
singulor moleculor componenl responsible ror thes.e fontostic
effecls . As wirh Ayohuosco, it wems likely Ihol odmixlures 01$0
ployed an imporlon! role in rhe monifestalon of Ihes. visionory
experiences. In this inlerasling hesis, rhe oulhors m.nlionad 01
leosl one plonr source for DMT which wos known lo be ossocioled
wilh Ihe Somo/Hoomo ceramony, Desmodivm gonganticum OC.
Parhops mora relevont moy be the menlion of bors.om, on odmix
ture wh ich wos curiously idenlifed os pomegronole in rhe lext.
Bonom wos olso refarred lo os o type of gro55, rhough in this
conlexl irs lilarol ond melophoricol meonings ore nol enrirely
cleor. Howover. severol .pecia. of Pho/ari. (found Ihroughoullho
worldJ ore known lo conroin significonr concentrolions of DMT
ond olher mOlhyloled Iryplomine. [Culvenor .1 ./. \ 964) . If ex
trocls from gron such os Pho/oris orvndinoceo were combined
wilh Ihe crushed or axtrocted s.eeds of P. horm%, Ihen on affico
cious bevaroge phormocologicolly similor lo Ayohuosco would
carloinly be obroined .
Through O similor piay between andogenous indoles, il has bean
during sJeep to promole Ihe visuol phenomenon of dreom slaap
[Collowoy 1988). ond o rece nI 'Iudy ho. le nI samo crodence lo
Ihi. ideo [Airo"inen el o/. 1993). According lo Ihe hypolha.i.,
lovel. of andogenou. ~C. increose during .Ieep ond focililole Ihe
The Iryptominas' oclivilias thus promota tha visuol ond amotive
componenl. of droom . Anolhor rece nI .Iudy hahown Ihollhe
inlarpodunculor nueleu. (o .moll oroo in Iho bosal mid-broin) ond
il. conn.clion lo Ihe hobo nulo, oro .... nliol fo, REMS [Huon 0101.
1992J. When Ihese connoclion. were cul in Iha rol broin, REMS
disappeared or docreosad dromolicolly. Aulorodiogrophic ond
recePlor binding ,ludia. wilh ['H)pinolin. havo .hown Ihollh.
highesl offinity of Ihi. THIlC derivolivo wo. highly localizod in lhe
inle'pedunculor nueleu. [Airok.inon el o/. 1993). Thi. andogonou.
ven ion of tha Ayohuosca experienca, os it ralotes 'o dreoming,
could ba o rolionoJe lor Iha noturol occurranca ond function of
p'ych.daliC indolo, wilhin Ihe humon CNS. Furlher re ..orch In
this oreo is onlicipotad .
When comporing Iha risks ond uncertoinlias nherantlo olher
meons of spiriluol davelopmenl, ona moy bagin lo underslond ,ha
volue of phormocologicol mOlhod . Indigenou. peopla. use ond
hove used ,hase porticulor subslonces for !heir raliobility lo lem-
InlegronOfl. jovrnol Jo,. ""rod-_vine pIorlll 0fI<I evlrv,., rw. 5/1995 7
pororily induce o visionory slole wilhin o wid. morgn of sofety.
The Iradilianal use 01 Ayahuaooa and alhor p.ychedelic .u~
slonces provide on individuollemporory occes.s 10 sloles of consei
ousness which ore normolly Inoc:cessible during Ihe woking slole.
In conclusion
Though Ihe seareh oanlinua. Ior somo univ.rsolly occepled and
ono1ytiooUy objectiv8 morkers lo diHerenliole betwHn 'us' OM
"Mm', in larms of mentol disorders, such lesls hove nol been de
voloped ond Iho doci.ion of sanlly .,ill remain. in ,he wolllrained
.ubjoclivily al Iho p'ychia,ri.,. In ,ho ruMo, moro .pocilicily in
onoiytiool procedures moy ollow us lo $N differences in groups of
individuols, Ihovgh Ihis is often olreody opporenl lo even ,he unlroi
ned eya. Since rhe role of psychosis increases proportionolly wilh
papulalion donsily, wo may do woll sludy ,ho habi'. al obcriginal
socielies which ahen lock equivolenl words far our concepts 01
depression, poronoio, hollucinolions ond suicide.
Since olh.r animols ha .... ye! lo speak 01 ony dosoge lo dale, such
research will 1011 u. liHIe abaullho prolound oHec'. al,hi. uniqua
closs of compounds fa, which human cuhures hove olready d..
monslrafed sofety ond volue, Indeed, Ihe cognitive effects enge"..
de red by o psychedelic subslonce moy be choraclerisric only lo
our own spedes,
It seems rhol o noturol funcl ion of psychedelic subslonces, wherher
endogenously produced or exogenou~y admislered, moy be lo
restare o sens.e of inner bolonce in the mind, Tha need lo enler
ohored "0'0. al mind i. undorscored by Iho locllhal .leop depri
votion quickly fesults in psycholic symploms ond il is nol unlikely
thol on ofhel sleep/dreoming mechonism could monitesl as cero
'ain Iypo. al p.ycho.... The cammanalily of Iho p.ych.delic
oxpori.nco indicalo. Ihal ,ho hoahhy mind ha. a oapacily and
lunclian lar .uch Slolo. likowise, Iho willrul induclion al a p.ych ..
delic slole is only on exlension of our noturol curiasity. Fat some
socielies il remoins on elfeclive technique lo occa" Ihe very
essence of Ihe soul. This powetful ond fundamental nslincl has
opporenlly nol diminished Ihrough feor or politicol repression, II
romoin. a Iranlior la be rully oxplarod ond devolopad by aur
Weslern cullures.
Acknowledgomonl.
Ma., al aur recenl wark ha. beon .upparl.d by granl. Iram Iho
Acadomy 01 Finland IModical Rosearch Cauneilj ond Iho Finni.h
Cullural Foundalian. Thanu la Anila Hommila Ior proal roading
the texl ond David POle for revie~ing In. monuseripl,
Nalo.
In Ihis contexl, consciousness refeu lo o stole of owofene" in
which ano i. awako ar able la unde,,'and whal i. happening,
ond chorocle,ized by o condilion which h05 or .shows on
8
understonding of ,he Mlf, on.'s surroundlngs ond Ih.
surroundings of olhers,
2 The lerm 'p.ychedolic' a. used in Ihi. article, rolo .. la Ihe
psychoocliv8 eHeclS chorocterislic of such dos.sk compounds
a. DMT, p.ilocibin/p.iIOCin, mo.calin. ar LSD In human . Tho
adjoclivo 'p.yehedolic', lilorally 'mind manife"ing' [O.mund
1957) or 'soul rovoaling' [PolO 1981), ,,ill can.id.rod la be
Ihe mo.1 appropria'. lorm 'o doscribe ,hi. eHocl [JaHe 1990).
legol 'erminology nol wilhslonding, Ihe lerm 'hollucinogenic' js
nol considered oppropriole, since 'fue hollvc.inolions ore
s.eldom ,he moin eHecl al o psychedelic subslonce .
3 1I .hauld be no,ed Ihal ,ho lorm 'hallucinalion', li'orally 'la
wonder in mind', is ohen defined os o sensory impression wilh
no bosis in eXlernol slimulolion . II is o subjective chorocterizoli
on u~d by rhose nol hoving ,he experience. Dreoms ohen Jock
such exlernol slimulOlon, bul ore s.eldom referrad lo os hall..,..
cinarions, even Ihough Ihay certoinly hJlfWlhe fequiremenls lO
be d.linod a ueh .
A Monoomine oxidase jMAO) jI a milochondriol enzyme rhol
oxid izes biogenic omines, ond similar exogenous chemicols, lo
deoclivo'ed watersoluble forms which con be el iminoled ffom
,he CNS. Inhibilion of Ihis enzyme results in on increose of
Ihes.e omines 01 ,hei, sile 01 oclion and a pOlenlolian of ,hei,
.Hects rhrough unchecked pasl.synap'ic aclivity. Some
anlidepro.sonl' wark by inhibiling MAO, and many af ,ho
oromolic tkorbolines ore good IV\AO inhi bitou, Inhibilors of
MAOA con ncreose concenlrolians of 5HT ond
noradrenoline, while inhibilors of MAO-B hove bHn used lo
ncreose levels of dopomine in polienls wilh Potkinson's
disoa... largo dose. al ma.1 MAO inhibilars will avorwholm
bo,h A and 8 lorm. al Iho onzymo.
References
Adachi, J.; Mizui, Y.; Naila, T.; Ogawa, Y.; UOlani, Y. and
Ninomiyo,l.
1991 . "Idonlificolian al 10lrahydro-Ikarbclino-J-corbcxylic ocid in
lood,'uH., humon urino and human milk: J. Nutr. 112:646-652 .
Airok.inon, M.M.; Ca llowoy, J.e.; Nykvi." P. ; Raga, L.; Kari. E.
and Gyn'hor, J.
1993. "Binding Silo. al ['H)Pinaline:
In: Toni'au, Y.; Arondl, J. and Pvol, P. [Ed. ) M.lolonin and Iho
. Pinoa/ G/and- Fram Basic Seieneo /o Clinica/ Applicolian. Am"or
dom, EI.ivior Seionco Publioalian B.V. pp. 83-86
Airok.inon, M.M.; Ho, B. T.; An, R. and Toylar, D.
1978 a. "Majar pharmacologieal effocl. al 6-melhaxy-lolrahydro
Il-carbalino, a drug olevaling Iho lissuo 5-hydraxylryplamlno
lovol." Arznoimi/lelFarsch. 28:42-46.
Airoluin.n. M.M.; Huang. J.T.; Ho. BJ.; TOylOf. D. ord Mdsoac. W .M. WyoH. R.J.

1978b. "Th. uplok. of 6-m.lho.y-1.2.3.4.1.'rohydrC>jl-co,ba1in. 1974. "Tryplolino" Formolion from Iryplominos ond 5MTHF by

ond il, .H.cl on 5hydro'y1ryplomin. uplok. ond r.1.ose in b100d humon plo'el."." Areh 01 G.n. P,yehio/ry. 31 :862-867.

plol.I."." Ac/. Phormocol. Toxico/. 43:375-380.

Borker. S.A. ; Monti. J.A. ond Chri,'ion. S.l.

Airoksinen, M .M. ond Kari, l. 1981 . "N.N-Dimelhyllryptomine: AA ondagenous hallucinagen." In:

19810. "!>Corbalin.,. p'ychOOCliv. campound, in Ih. mommo Inrerna/ional R.view al Neurobio/agy. Vol. 22; Acodemic Pr.... loe.

lion body. Porl 1: Occurr.nce. origin ond m.'oboli,m ." M.dicol

Biolagy 59:21 -34 Beck. O . ond Foull. K.F.

1986. "Conaentrolions of Ihe enontiome" of 5-hydro.y1ryploline in

Airo.,in.n. M.M. ond Kori. 1. mommolion urine: Implico'ions for in vivo biosynlh.sis.... Biocm.m.
1981 b . "l-Corbolines, psychooctjve compounds in rhe mommo Phormaeol. 35:2636-2639.

lion body. Por! 2: EHec..." Medicol 8iology 59 : 190-211.

Bick.l. P.; Dinrieh. A. ond Schoeph. J.

Airoksinen, M .M. ond Mclsooc, W .M. 1976. "Allered ,tore, of oon>ciousn... induoed by N.N-dime

1968 . "Indoloolkylomino, ond bohovior ." Ann. M.d. Exp. Fonn . Ihyltryptomino." Pharmakop'ych. N.urop,ychophormokol. 9:220-225.

46 :367381 .
Bculton. A.A.

A iroksinen, M .M. ; Mhonan, M .; Tuomislo, L.; Pauro, P. ond 1971. "Bioch.miooll.search in schizophronio: NO/lJfe 231 :22-28.

Erikuo n. c.J.P.

1983. "T.'rohydro-{korbalino" Effecl on olcohol inloke in ro ... "

Bo,in, T.R .; Foull. K.F.

Phormocol.. Bioch.m & B.hov. 18 :525529.
Airoksinen, M.M.; Soinio, EA .; leppluoto, ond Kori, 1.
1984. "6-Melho.y'.'rohydro-ll-carbolino (pinoline): Effecl' on
plo,mo r.nin oc~viry ond oIdo,'.ron THS. LH ond Hondorphin
l.v.l, in rOl': Aclo Endocrinolagio 107:525-530.
1989 . "Indole d.rivolizolion procedures far electron capture
n890liv8 chemcol ionizolion mou spectromelry: Identificaran of
l.melhyI-1.2.3 .41.'rohydro-Xolbolin. in rOl broln ond lung:
Biomed Enviran. Ma... Spec/rom. 18 :247252.
Bo,in. T.R.; Fovll. K.F. ond Boreho,. J.

1988 . "Hormon in olcoholic b.velogo" Pholmocolagical ond

Airoksinen, M .M .; Svensk, H.; Tuomislo, J. ond Komuloinen, H. lo.ieolagicol implicolion . ... Alcohol. Cln . Exp. Ro,. 12 :679-682.

1980. "T.'rohydro.korbolin., ond carro'ponding Iryplomine,. In

by humon blood
vivo inhibirian of serclonin ond dopomine uploke Broe",up. c.; Niol",n. M. ond Ol,en . C.E .

plol.I.,,". AClo Phormocol. Toxicol. 46 :308313 .

1980. "Urinory ond broin Xolbolin&-3-corba'yloIO' o, polonl

inhibilors of broin benzodiozepine receptors ," Proc. NoH. Acod.

Alond.r. O.R.
Sei. 77 :22882292 .
1762. In: Inobrionlio (ocodemic di,serlOlion und.r C. Linno.u,).

Univ."iry of Uppsolo. pp. 10-11.

Brun. G .G. ond Himwich. H.E.
1962. "Indole metabalil8! in schizophrenic polients. Urinory
Apud. JA
.xcr.lion ." Arch. Gen . P'yehio/ry 6:324-328 .
1991. "The 5-HT, rec.plor in broin: Rec.nl biochomicol ond

molecular biologicol developments ond new pers~clives in ils Buckholtz. N.S.

I.gulolion: Phormocol. R.,. 23:217232. 1980. "Neurobiolagy of letrohydlo-{korbolino,: lile Sei.

27:893903 .

Anlun. F.; Burn.n. G .B.; Coopero A.J.; Doly. R.J_: Smy1hio,. J.R.

ond Zeoll.y. A.K.

Buckhollz. N.S. ond Baggon. W.O.

1971 . "ThH.cl, of L-melhionino (wilhoul MAOI) in schizophre

1977. "Monoomine oxidose in broin ond livor produced by

nio: J. P'ychio/ric. Ro,. 18 :6366.

p<orbolinss: Slfuclure-oclivity relolionships ond substrole specificity."
Bioch.m. Phormocol. 26: 1991 -1996.
A I,od. J.

1961. "Enzymolic formol ion 01 p'ycholomim.lic melobalil., from

Bumpu,. F.M . ond PoS. I.H.
normolly occurring compound,: Seienco 134:343.
1955. "Serolonin ond ils melhyloted derivativas in humon urin.,
1962. "Thnzymolie N-melhylOlion of serolonin ond olher
1. Biol. ehem . 212:111-1 16.
omin.,: J .Phormoeol. Exp. Th.rop. 138 :28-33.

BO$zormnyi. A . ond Szoro, S.

Borcho,. J.D .; Elliol. G .R.; DoAmorol. J.; Erd.,lyi. E.; OConnor.
1958. "Dim.,hyl'ryp'omino p.riments with p,ychOlics." J. Mon/.
S.; Bowd.n. M.; Brodi. H.K.H.; B.'gor. P.A.; Renson. l. ond
Sci. 104:445-453 .

9
Collowoy, le.
Fobing , H.O . ond Howkin., J.R.

1988. "A proposed mechonl.m for the vi. ion. of dreom .Ieep."
1956. "Introvenou. bufolenin. njec.ion In th. humon being."

Madicol Hypolhe ..., 26: 119124.

Science 123:886-887.

Forrel, G . ond Mclsooc, W .M.

Collowoy, le.
1961 . "Adrenog lomerulo.ropin" (Iener.o .he edilor). Arch. Bio

1993, Tryptomine. p<orboline. ond you . MAPS New./ener of Ihe

chem. 94:543544.

Mu/lidisciplinory Associolion of P.ychedelic Sludie. 412) :30-32.

Fischmon, L e.

Collowoy, J.C; Morimoto, H.; Gynther, J.; Airo"'inen, M.M. ond

1983. "Or80ms, hollucinogenic drug slolo. ond schizophrenia : o psy

W illiom., P.G .
chologicol ond bialogicol comporisoo." ScJ,izophren. BuN. 9:73-94.

1991 . "Synthe.i. of ('H]pinoline, ond endogenou. tetrohydro-~

carboline." J. Lob.lled Compel. Rodiophorm. 31(5):355364.

Fish, M. S.; Johnson, N. M. ond Horning, E. C.

1955. Pip.odenio olkoloid . Indole ba .... of P. peregrino (l.)

Collowoy, J.e.; Gynther, J.; Poso, A.; Vepsloinen, l ond Airok.i

B.n.h. ond 'elo d .peci ." J. Am. Chem. Soco 77:58925895 .
nen, M. M.

1994. "The Pictet.Sprengler reoction ond biogenic tryptomines:

Flon.ry, O. S. ond Schwor'z , M.
Formo.ion of .e.rohydro1l<orboline. o, phy.iolog icol pH ."
1989 . In : Horno ond Hormalin.; Uni.,."ity 01 ColilOl"n;o Publicori
J. Helerocyclic Chem. 31 :43 1-435 . ons: Neor Eo,t.rn Sludi." vol. 21 , S.,keley, Unlversity of Colifor.
nio P,.... pp . 32, 36, 40 & 41 .
Chri.tion, S.T.; Horrison, R.; Quoyle. E. ; Pagel, J. ond Mon.i .
19n. "Th. in vilro id.n.ifica.ion of dime.hyhryp.omine (OMn in Friedhofl, AJ .; Schweilzer, J.W . ond Miller, l
rnommolian broin OM ils chorocrerizotion 01 o po"ible endog.. 1972 . "Bio.yn.he.i. of meseoline ond N-oc.tylmescoline by
nov. neuroregulo.ory ogenl. " Biochem. Mad. 18 : 164 183 . mommolion liver." Nolure 237 :454-455.

Cohen, S.M.; Nichol., A; Wyon, R.l; el 01. Friedhofl, Al ond Von Winkl., E.
1974 . "The adm inistrarian of melhionine lo chronic sch izophrenic 1962. "The choroclerislics of on omine found in ,he urne of
po'ien's: A review of 'en ,.udie . " Bio/. P'ychiolry 8:209225. s.chizophrenic polienls." journol af Nervous ofld MefllOl Ds&ose
135:550-555.

Corben, l .; Chri,'ion, S.T.; Morin, R.O.; Bening.on, F. ond Smy

hie., J.R.
G...n.r, P.K. ond Pog., I.H .

1978 . "Hollucinogenic N-methyloted indoleolkylomine. in .he

1962 . "B.hoviorol efleclS of 5m.thoxy-N,N-<lime.hylrryp.omine,

cerebro.pinol Auid of p.ychio'ric ond control populotion ." 8ril. J.

o.h.r tryplomine. ond LSO ." Am. ). of Phy>iol. 203 : 167 172.
p, ychiolry 132: 139144.

G illin, J.e. ; Koplon, J.; Stillmon, R. ond Wyon, RJ.

Croo"', P.A.; Godin, e.S.; Nwosu, e.; Ansher, S. ond Jocoby. W .B.
1976. "The psychedelic model of schizophrenio : Th. ca... of

1986. "Reevo luo'ion of Ihe producl. of Iryplomine co.olyzed by

N,N-<lim hylrryp.omin." Am. ). P, ychiolry 133 :203208 .

robbilliver N.m hyhron.fero ... ." Biochem . Phormoco/. 35 : 1600

1603 .
Glennon, R. A.

1990. "Do clo..ico l hollucinog.n. oc! os 5HT2 ogonis.s or

Culvenor, C. C. J.; Bon, R. O. ond Smith, l . W .

onlogoni.l.i" N.urop,ychophormocofogy 3:509517.

1964. "The occur.nce 01 indoleolkylomin. olkoloid. in Pholori.

rvberoso l. ond P. orundinoceo l." Au,l. J. Chem . 17: 130 1 1304.

Guchhoil, R.B.

1976. "Biogenesis of 5.m.lhoxy-N,N-<limelhyl.ryptom in. In

Oeligon i. , AV.; Pierce, PA ond Peroulko, SJ.

human pineol glond." ). Neurochem. 26 : 187190.

1991 . "Oiflerentiol inl.roclion. of dimethyl'ryp.omin. IOMn wi.h

5HT, ond 5.HT, receptors." Biochem. Phormoco/. 41: 17391744.

Gunlh.r, R. T.

1934. In : The Greek Herbol 01 Dioscoricle,. Oxford University Pr.".

O.m.nt, W.

1960. "The eflect. 01 dreom deprivollon." Science 131: 1705 .

Gynther, l; lopinjoki, S.P.; Airokslnen, M.M. ond Peuro, P.

1707.
1986. "Decorboxylolion of 1,2,3 , 4 t.lrohydro-~o rbolin ... l.

corboxylic ocids in broin homog.not. ond cololysis by pyridoxo~

Elliot, G .R .ond Holmon, R.B.

5'.pho.phole." 8iochem. Phormocof. 35 :2671 2675 .

19n. "Tryptolines o. potentiol modulo toro of ... rotonorgic

funclion." In : Usdin, E. (Ed.] Neuroregulotors ond p'ychotric

Gobel, H.

disorders. O xford Univ.rsity Press. pp 220-226.

183B . "b.r d ie G.winnung ein.r rolhen Forb.loffes ou. d.n

lO

Somen der PAonze Pegonum Hormolo ." Oing/ers Po/ytechni,cho.

T. W .; Nies, A. S. ond Toylor, P. IEds.) GocxJmon on Gi/mon'.
Journo/, S. 373376.
The Phormoco/ogico/ Bo,i, 01 Theropeuffcs, New York, Pergomon
Press. chapler 22. pp. 553557.
Ha, B.T.

1972. "Monoomine oxido.. Inhibilo"." ). I'harm. Sci. 61 :821837.

Kori, l.; Alroksinen, M.M.; Gynlher, J. ond Huhtlkongos, A.

1983. "Moss speclromelric idenlilicolion 01 6.melhaxy-l ,2,3,4

Ha, B.T.; Toylor, D.; Askew, W .E. ond Mclsooc, W.M.
1.lrohydro-iXorboline in pineol glond: I/ec. Oev. Moss. Spec

19720. 'Effecl, 01 6-melhoxytelrohydro-jkorboline on Ihe regio

/rom., Me. on Environ. l/es. 8: 19-24.

nal ond sube.llulo( dislribulion of serofonin in mause ond robbit

broin,." Lfe Sei. 11 :493502. Kellar, K.J .; Elliol, G.R .; Holmon, R.B.; Borchos, J.D. ond Verinko

Donnellis, J.

Ha, B.T.; Toylor, D.; Wolker, K. ond Mcl..oc, W .M.

1976. "Tryploline inhibilion 01 ..rolonin uploke in rol lorobroln

1972b. "The mode 01 oclion 01 6-melho.y.t.trohydro-iXorbaline

homogenole: ). Phormoco/. Exp . Ther. 198 :619-625 .

on broin serolonin: Con. ). Biochem. 51 :482-485.

Ku.hl, FA; Ormand, R.E. ond VondenH~';'el, W J.A.

Holm'l.dl, B.
1966. "Occurrence of 3,4-0imelhoxyphenylocelic ocid in urino 01

1982. "Belocorboline. ond 1.lrohydroisoqulnolin.s: hi,loricol ond

normol ond schizophrenic individuols."
thnophormocological background: In Bloom, F.; Borcho" J.; No/ure 21 1:606-608 .
Sondlor, M. ond Usdin, E. IEd,.) Be/acerbo/in., ond to/rohydroiso
quinohn., Iproce.ding' al o worbhop h.ld 01 Ih. Solk In.lilule, lo KOrkJ<Oinen, J.; Riiisainen, M.; Noukkorinen, H.; Spoov, J. ond Riman, R.

Jallo; California; Docember 12 ond 13, 19811. N.w York, Alan 1988. "Urinory excre/ion of free bulolenin by p,ychiolric poli

R. li... pp. 313. enls." Bio/. Psychio/'Y 24:441-446.

Holm.tedt, B. and Undgr.n, J. .e.

leino, M. ond Airaksinen, M.M.

1979 "Ch.micol con.tituenls and phormocalogy of Soulh Ameri

1985. "Melhoxyindoles In relino: Me. Bio/. 63:160-169 .

can .nulls: In: Efron, D. H.; Holm.l.dt, 8. ond Klin., N. S. IEds.j

E/hnopharmc%gic Seorch lor P.ychooc/ive Orug' [proc.eding. 01

Uoyd, EJ. ond Androws, P.R.
o .ympo.ium h.ld In San Francisco, Jon. 28-30, 1967}. New 1986. "A common slruclurol modal for central nervous system
York, Rov.n Pr.... pp. 339-373. drugs ond Iheir receploro: ). M..a. Chem. 29:453-462 .

Hochstein ond Parodies luna, L.E. ond Amoringo, P.C.

1957. "Alkoloid. 01 Boni,to,;o coop; ond Pro.tonio omozonicum: 1991 . Ayohuosco Visions. Berkeloy: Norlh Allan~c Books.

). Am. Ch.m. Soc. 79 :5735.

Mondell, AJ. ond Morgon, M.

Hryhorczuk, l.M,; Roiney, J.M.; F/ohom, e.E. ond Novok, E.A.

1971. "Indol.lelhyl}omine N.melhyhron.l.rose In humon broin."

1986. "A n.w m.tobolic polhwoy Ior N,N-dim.lhyhryptomine:

Noture 230:8587.

Bio/. P.ychio/'Y 21 :84.

Mol.ubaro, K.; Collins, M.A. ond Neohay, EJ.

H,u, L.l. ond Mondell, AJ.

19920: Mono-N-melhylolion 01 1,2,3,4lelrohydro-lXorbolines In

1975 . "Enzymolic lormolion 01 lelrohydro-lXorboline. Irom

broin cyto>ol: Absence 01 indole molhylotlon: ). 01 Neurochem.

Iryplomin. ond 5.me/hyh.lrohydrololic ocid in rol froclions:

59(2) :505.510.

Regional ond .ubcellulor di.lribulion: J. Neurochem. 24:631

636. MOI.ubaro, K.; Fukushimo, S.; Akone, A.; Horno, K. ond Fukul, Y.

1986. "T.lrohydro-lXorbolines in humane urine ond rol broin- No

Huon, F.; Eckenrode, r.e. ond Murrou .vidence 01 lormolion by olcohol drinking : Alcoho/ & Alcoho/i,m

1992. 'Hobenulo ond Iholmu. celllron.plonts re.lore no/mol 32:368372 .

Ieep behovioro disrupled by den .. rvolion 01 Ih.. inl.. rpedunculor

nuci.u . " J. N .. urOJCi. 12:32823290. Molsubaro, K.; Neol..y, EJ. ond Collins, M.A.

1992b. ' Novel S-odeno.ylmelhionino-dep.nd.nl indol ... ~elh)'

Jocob., B.L. ond Trul>on, M.E.

lotian af JJ-<;orbolines in broin porliculote froctians. J. Neuro
1979. 'Dreom" hollucinOlion, ond p.ycho'i5- The ,erOlonin
chem. 59:511-518.
cannoclion: Trend, in Neuroscience 21111:276-280.

Mourizi. C.
Joffe, J.H.
1984. "A mechoni,m 01 monia 000 Ihe cheml.,ry 01 droom.:
1990. "Drug addiclion ond drug obu... In: Gillmon, A. G.; RolI,
A hYPolhesis: Sou/h.," Medico/ Journo/ 77 : 14911493.

11
May, T.; Rammelspaeher, H. and Pawlik, M. Narosimhaehori, N.; Boumonn, P.; Pak, H.S., 01 al.
1991. "['H]Harman binding experimonls . 1: A rovo"iblo and 1974. "Gas chrama!ogrophic-moss speelrame!rie idenlinealian 01
Mtlecti.... e rodioligand far monoom;ne oxido~e A in the CNS of 1he urinory bufolenin and dimelhyllryplomine in drug free ehranic
rol." J. Nourachem. 56 :490499 . schizophrenie pOlienl'." Biol. P'yehiolry 8 :293370.

Melsaac, W .M.
Nichol" D. E.; Oberlender. R. A. ond MeKenna, D. J.
1961 . "Farmalian al l .melhyl-6melha xy1 , 2 ,3 ,4'lelrahydro-2
1991. "Sloreachomieol a,peels 01 hollueinogens." In: Walson, R.
corboline under physiolo9icol condilions." Biochem. Biophys. IEd.] Biochemi"ry and Phy'iology 01 Sub,'ane. Abu,e; val. 111.
Aela 52 :607-609. Baco Rolon, CRC Pro". pp. 139.

Mclsaac, W .M.; Taylar, D.; Walker, K.E . and Ha, B.T.
1972 . "6-Melhoxy-t8ImhydrQ-Jl<:orboline- o s.erolonin elevolor."
j . Neuraehem. 19 : 12031206 .
McKenna. DJ .; Ropke. D.B.; la. l. and Peraulka. SJ.
1990. "DHerenliol inleroclions of indoleolkylamines wilh
5hydroxylryptomine re~plor subtypes ." Neurophormocol.
29 : 193 19B .
McKonna, DJ . and Tawors. G.H .N .
54171175.
O,mond, H.
1957. "A re.... iew of Ihe clinicol eHecls of psychOlomimelie
agenl'," Annal, ollhe New York Aeocl.my 01 Seionco, 66 :418
434.
Oon. M.e.H.; Murray. R.M.; Radnighl. R.; Murphy. M.P. and
Borloy. J.l.T.
1977. "Factors oHecling Ihe urinory excrelion of endogenousfy
formed di melhyllryplomine in normal subjecls." Psychopharmaco/.

1984. "BiachomiSlry and pharmaealagy al Iryplamines and beta

corbotines: A minireview," J. Psychooclve Drugs 16:347. Oomand, H. ond Smythio,. J.R.

1952. "Sehizophrenia: A now approach." J. ....... nl. Sei. 98 :309315 .

MeKonna, DJ .; Tawers, G .H.N . and AbaN, F.

1984. "Monoomine oxidose inhibitors in Soulh American halluci POle. D.W.

nogenic planl": Tryptomines ond belo-carboline consliluenls of 1981. Personal communicolion.

Ayahuosca." J. Elhnopharmocal. 10: 195.
Poa, e.. Modina , J.H .; Novas, M.l.; Poladini, A.e. and D.
Mcl&od, W.R. and SilOrom, 8 .R.
Roberlis, E.

1985. "Bufolonino reoonsidered." Acb P'ychiotry Scond. 72:447450.

1986 . "Isololion ond identificalion in bovine cerebral corte x of rt

burylXorbaline-3<orboxylole, o polen! benzodiozepine binding

Moller, E. and Guha, S.R .

inhibilar." Prac. Na/l. Acad. Sei. 83 :4952-4956 .
1979. "'nhibition poMarn of monoomine oxido!oe in sub-froclions
of ral broin milochondrio in !he pre)08nce of some selecti.... e inhibi Peura, P.; Koti, 1. and Airaluinen, M .M .
lors." Biochem. Pharmoeal. 2B : 11351137 . 1980. "Idenlifiealian by ..Ioelive ion manilorlng al l.melhyl.
1. 2 ,3,4lelrahydrall<arbafine in humon plalelel' ond plo,mo ol1er
Meyer, J.S.: Buekhollz, N .S. ond Boggan, W.O . elhanal inlake: Biomed. Mo ... Spoc/rom . 7:553555.
1978. "Serolonergic slimulolion af piluilory<ldrenal ocli .... iry in the
mou.... Neuropharmocol. 26 :312 324.
Peura, P.; Jahnson, J.V.; Ya,'. R.A. ond Faull, K.F.
1989. "Concentrarians 01 tryPloline ond melhlryploline in rol
Meyers, R.D. ond Molchior, e.l.
brain ." J. Neurochem. 52:847852.
1977. "Differenliol oelion, al volunlary olcahal inlake al lelrahy

droisoquinalines ar o J}-<:arbaline inlused chranically i n ,he ....enld Pollin, W .; Cardon, P.V. ond Ke/y. S.S.
cal 01 Ihe rol.' Phormoeo/. Biachem. Behov. 7 :381 392 . 1961 . "Effecls alamina ocid feedings in schizophrenic polien!s
Iroolod wilh iproniozid." Scione. 133: 104-1 05.
Morgan, M. and Mondell, A.J.

1969 . "lndolelelhyl)omino N.melhyltronslero,e in Ihe broin."

P,ehoidl, G.R.; Berlel, H.H.; Spoide, l. ond Himmwich, H.E.
Scienee 165:492-493.
1966. "Voriotions of uri nary creoHnine ond ils carrelotion lo
excrelion of indole meloboliles in mentol palienls," Clin. Chem.
Noranjo, e.
Ac/o 13 :228.
1967. "Psycholropic praperlios al hormola olkaloid ." In : Elron,

D. H., Helm,'ed" 8. and Kline N . S. IEd,.] E'hnophormocologic Riceberg, l.J . ond Von Vunokis, H.

Seorch lar Psyehoaclive Drugs Ipraceedings 01 o 'ympo,ium held 1978. "Delerminalian al N,N-dimelhylindaleolkylamines in plosma,

in Son Froneisco, Jan. 28-30, 1967). New York, Roven Pre". blood and urine extraels by radioimmunoassoy and high pressure
pp. 385-391. liquid chromalogrophy. ). Phormoool. Exp. Th.r. 206: 158166.

12
Rommel'pocher, H.; Bode, B.; Cooper, H. ond Kossmehl, O. Shulgin, A. T.

1976. "Inhibilion 01 uploke 01 serolonin by Iryploline." 1976. "Proliles 01 p,ychedelic drugs. 1. DMT." Journol 01 P'ych.

Noun yn.Schmiedeberg', Arch . Phormocol. 292 :93 .95. de/ic Drugs 8: 167 168.

Rommel'pocher, H.; Domm, H.; Slroull, S. ond Schmidl, G. 1977. "Proliles 01 psychedelic drugs. 4. Hormoline."

1984 . "Elhonol induces on "crease of horman in the broin ond Journol 01 Psychedolic Drug' 9:7980.

urine of rols." Nounyn.5chmiedebefg '$ Arch. PhormocoJ.

327:107113.
1981 "Proliles of psychedelic drug'. 11. Bulolenine."

Rommel'pocher, H.; Moy, T.; ond Su,ilo, R. Journol 01 Psychede/ic Drugs 13 :389.

1992 . "~orbolines ond letrohydroiwquinolines: Dereclion ond

lunclion in mommol,." Plonro M.d. 57 :B592. Shulgin, A.T. ond Shulgin, A.

1991. Pihko/: A Chemicol Lave Story. Berkeley, Tronslorm Pr....

Rommel'pocher, H.; Nick, M. ond Brning, G. pp. 6 14- 16.

1984 . "An inlermediole product in rhe jn vitra synlhesis of horman

ond olher Xo rboline,." Nounyn.Schmiedeborg ', Arch. Phormo Silorom, B.R.; Blockmon, G .l. ; Mcleod, W.R. ond Voughon, G.N.

col. 325:R76. 19B3 . "The ionpoir extroction, purificarion, ond liquid chromoto
g raphi c analysis af indoleolkylamines in human urine: Anal.
Rommelspocher , H .; S'rouss, S. ond undemonn, J. Bioch.m . 128: 11 20.
1980. " Excrelion of 181rohydrohormone ond hormone inlo Ihe
urin. 01 mon ond rol after O load w ilh erhonol. ... FEBS (aUe" Smythies, !.R .

109:209212. 1960. "Recenl Advonc.s in Ihe biochemislry 01 psychosis."

10ncol vol. 1 no . 7137, pp . 1287 1289.

Rosengorlen, H. ond FriedhoH, AJ.

1976 ... A review aflhe recenl sludies of Ih. biosynlhesis ond Slrossman, R. J.

excrelion of hollucinogens formad by melhylolion 01 neurolronsmil 1992. "Human hallucinogenic inferaetians with drugs off&cling

le" ond reloled ,ubstonces." Schizophr . Bu/l. 2 :90105 . s.6roloergic neurolransmission." Neuropsychaphormaco/.

7(3J:24 1243.
Rosengorlen, H.; Meller, E. ond FriedhoH, A.).
1975 . "Synlhesis of letrohydr~orbolines frcm indoleomines .... io Slromberg, V.l.
enzymolic formotion of formoldehyde from 5-merhyltetrohydrofolic 1954. "The iwlalian of bufotenine fram Pip,oden;o fMregr;na."
ocid." Biochem , Phormocol. 24:17591762 . 1. Am. Ch.m. Soco 76: 1707,

Rosengorlen, H. ; Meller, E. ond Friedholf, AJ. Suslo, R. and Rammelspocher , H .

1976 . "Possible source of error in sludies of Ihe enzymolic formal; 1988. "Formolion 01 l.melhy4Xorbolines in ro" from Iheir po..i

on 01 dmelhyllryplomine." ). Psychiolric Re" 13:2330. ble corboxylic ocd precur",r." Nounyn.$chmi.deborg', Arch.

Phormocol. 337:566-571.

Risnen, M.
19840. "The presence 01 Iree ond eonjugoled bufolenin in normal
Szoro, S.

humon urine." lile Sei. 34:20412045.

1961. "Hollucinogenic eHects ond meloboli,m of Iryplomine

derivotives in mon."
Risnen, M. ond Krkkoinen, ).
Fed. Proc. Fed, Am. Soco E~p. Biol. 20:885888.
1979. "Mo.. Irogmenlogrophie quonlilolion of uricory N,N

d imerhyhryplomine ond bulorenin." 1. Chromologr. Biomed. Appl.

Tomos, T.S.; Buckhohz ,N.S, ond Zemp, ),W.

162:579584.
1979. "6-Melhoxy.I ,2,3,4.lelrohydro-lXorboline eHeels on r.linol

serolonin." Lilo Sci. 25: 1435 1442 .

Risnen, M.; Virkkunen, M.; Huttunen, M.O.; Furmon, B. ond

Krkkiiinen, J. Turner, W J. ond Merli" S.

1984b. "Increosed urinory excrelion of bufotenin by violen! offen 1959 . "Elfecls of some indol.olkylomines on mon." Arch. N.urol.

de" wilh poronoid symploms ond family violence." Loncel vol. 11 P,ychiolry 81: 12 1129.

No. 4B04 pp.700-70 1.

Tuomislo, l.; Airo"inen, M.M.; Peuro, P. ond Erlksson, CJ.P.

SonchezRomo" J,R. 1982. "Alcohol in Ihe rol: Increose, fallowing inlrocerebrov.nlri

1990 . "Review: Bonis!erine ond Porkinson's Diseose ." cular Irealmen' wi,h le'rohydro-~orbolines." Phormaco/. Biochem.
Clin . Neurophormoeol. 14:391402. Behov. 17:831836.

13

,.

Torro., e. M.; Ropko, D. B.; Chon, K.; McK.nno, D.; llogo'I.",

A. ond Schult.., R. E.

1991. "Snuff powd ... Irom pre-hi.ponic Son Podro de Alocomo:

Chomicol ond conl.xluol onoly.i . " Cvrr. Anlhro. 32:640-649.

V.rl, R.P.

1986 .A lil.... u.loining lunclion lar REM .Ioop: A Ih80ry." Nov

rosc. & Biobohov. Rov. 10:371-376.

Wokoboy.hl, K.; Ochiol, M.; Soilo, H.; hudo, M., Suwo, Y.;

Nogoo, M. ond Sugimuro, M.

1983. Pr....nco 01 l-molhy~ 1,2,3,4-telrohydro-jkarboline-3.

corboxylic ocid, O precursor of o mulogenic nitroso compound, in

soy sauco. Proc. Norl. Acod. $ei. USA. 80:29122916.

Wolk.r, R.W.; Mond.l, l.R.; KI.inmon, J.E.; Gillin, J.e.; Wyon,

RJ. ond Vond.nH.uvol, W J.A
1979. "'mproved selective Ion monitoring mo","spectromelric
ouoy far Ihe determinatian of N,N-dimelhyllryplomine in humo n
blood ulilizlng capillory column go. chromologrophy. J. Chromo

Iogr. Biochom. Appl. 162:539-546.

Wo..on, R.G.; Holmonn, A ond Rvck, e.

1978. "The Rood lo Eleusi.. Now Y",k, Horcourt Broce Jovonovich.

W lern, O.e. ond Ozburn, E.E.

1949. "Melhonol ond formoldohyd. in normal bocy li ..u ond

fluid . " UnilecJ 5101.. Noval Bul/olin 49:574-574.

Whol.y, W. H.; Govindochori, T. R.

1951. "Th. Piclo~Sponglor 'ynlhi. 01 I rohydroisoquinolin

ond reloted compounds," In: Orgonic Reoch"ons vol. 6. New York,

Whil.y. pp. 151191.

Wyon, RJ.; Erd.lyi, J.E.; DoAmorol, J,R.; Elliol, G.R.; R.n",n,

J, ond Borcho., J.D.

1975. "T ryplolino formelion by o pr.porolion from broln wilh

5-melhyltelrohydrofolic ocid ond Iryplamino. Scionoe 187:853-855.

Wyon, RJ.

1985. "Tho dopomlne hypolhe.is: Voriolion. on O Ih.m.

In: Conceo, R. ond Doon, S. R. [Ed. ] Re.eorch in Ihe Schizophr..

nie Disord.... Th. Stanloy R. Deon L.elvro. vol. 11. N.w York,

Spoclrum Publicollon, pp. 225247.

Youdim, M.B.H. ond A.hkenozl, R.

1982. "Rogulalion 01 5HT caloboli.m. In: Ha, B.T.; Schoolor,

J.e. ond Usdin, E. [Ed. ] Sero/onin in Bio/ogieol P,ychiotry.

Now York, Rov.n Pro.., pp. 35-60.

14
VI

Psychopharmacology (1994) .:....

Psychopharmacology
Springer-Verlag 1994

Platelet serotonin uptake sites increased in drinkers of aya huasca

James C Calla way l.2", Mauno M AJrakslnell ' , Dennls J McKenna J , Glacus S Brlto" Charles S Grob
'
I Deparlmenl of Pharmacology and Toxicology, UniversilY of Kuopio, POB 1627, FIN70211 Kuopio, Finland
l Deparlmenl of Pharmaceulical Chemistry, Universily of KUOplO, POB 1627, FIN70211 KUOplO, Finland
J BOlanieal Dimensions, PO Box 807, Occidenlal, CA 95465, USA
CGnlro De Esludos Mdicos D. Uniao Do Vegelal, Caixa Posl.171505, 05020990 Sao Paulo - SP, Brasil
1 Deparlmem of Psyehialry, Bldg. D-6, Harbor/UCLA Medieal CGnler, 1000 Wesl Carson Slrtel, TOrTance, California 90509, USA

Abstracl.- The binding of [3H)cilalopram 10 Ihe plalelel
5-hydroxylryplamine (5-H"!') Iransporter was measured
in a group of heallhy male drinkers of ayahuasca, a psy
choaclive subslance indigenous 10 Amazonia, and a
group heallhy male conlrols. An increased number of
binding siles (B m ,,) in Ihe plaleles of ayahuasca drinkers
was found, while Ihe dissocialion conSlanl (KJ re
mained Ihe same for bOlh groups. If indicalive of neuro
nal 5-HT uplake aClivilY, Ihese resulls would suggest a
decreased concenlralion of eXlracellular 5HT, or a re
sponse lO increased produclion and release of 5HT
Such changes in 5-HT synaplic aClivily. in Ihis case,
should nol be misinlerpreled as an indicalion of develop
ing Ileurological or psychialric illness.
Key words: fJ-Carbolines - Dimelhyltryplamine (DMT)
- Elhnopharmacology - Harmala alkaloids - Human
plalelels - 5-Hydroxytryplamine - Psychedelic
lntroduction
The religious use of psychedelic subslances (also known
as hallucinogens) predales wrillen hislory and survives
to Ihis day bOlh in pre- and posl-induslrial cullures. Lil
Ile is known aboul Ihe long lerm effecls from Ihis Iype of
drug use. We have recently begun a comprehensive in
vesligation of ayahuasca. since Ihe chemical nature of ils
aClive consliluenlS and Ihe manner of ils use are relevanl
10 conlemporary issues in neuro- and psychopharmacol
ogy.
Ayahuasca is a Quechua lerm meaning "vine of Ihe
soul", which applies bOlh lO Ihe beverage and 10 Ihe vine
Ballsleriopsis caapi, one of Ihe major planl componenls
from which ayahuasca is brewed. This " lea" is also
known as caapi, yage, hoasca, vegelal and daime. lis ori
gin is mylhological and il is slill widely used by bOlh ab-
Correspondence 10: Le. Ca11aw.y. Dcpl. of Ph.rm. Chem. FAX
1/35871 162456
original and indigenous Meslizo populations Ihroughoul
Ihe Amazon Basin.
Harmala alkaloids, primarily harmine (l-melhyl. 7
melhoxy'Jcarboline), are infused from Ihe pounded
woody portions of B. caapi and serve 10 reversibly inhib
il monoamine oxidase A (MAO-A) . Some 'Jcarbolines
can also inhibil Ihe neuronal uplake of 5HT (Ajraksinen
and Kari 1981). The harmala alkaloids are essenlially
devoid of psychedelic aClivily at doses obtained from Ihe
lea (1-3 mg!kg). Tremors. nySlagmus, vomiling and di
arrhea are the common fealUres somelimes simulta
neous ly achieved wilh higher doses of pure harmine or
harmaline (3-5 mg!kg. po; unpublished observalions).
N,N-Dimethyltryplamine (DMT), a second compo
nenl of Ihe lea, is obtained from Ihe lea ves of Psycholria
viridis. DMT is a potent and short aCling psychedelic
agent when smoked or injected, bul is orally inaclive due
10 ils rapid oxidalion by MAO-A. Togelher, the MAO-A
inhibilion by harmine wilh lhe oral psychoactivily of
DMT form the basis of this ancient rite (for excellenl re
views of ayahuasca see Luna and Amaringo 1991.and
011 1994).
We were inviled by Ihe Unio do Vegelal (UDY), a
syncrelic religious movemenl in Brazil, lO collecl blood
plalelels and other samples. The membership of this
group exceeds 5000 and includes several physicians and
olher heahhcare workers. Their general knowledge con
cerning Ihe bOlany, chemistry and pharmacology of ay
ahuasca is impressive . The aim of the present part of this
study was 10 delermine if chronic use of ayahuasca re
sulled in any measurable longterm changes in Ihe 5-HT
Iransporter.
Materials and methods
Thirteen hcaithy volunleers <a11 male members of Ihe UDV be
twccn 28 and 48 ycars of age; mean ageslandard deviation,
38.55.6 ye.cs,) who h.d drunk Ihe le 1 le,sl biweekly for 10
years or more, and len controls (<tll males aging from 21 to 45
yars; mean agestandard dcvialion. 33.27.9 ycars) who had
ncvcr drunk th e lea werc includcd. Al! subjecls wcre re sidcnls of

Ms. No. 503 Aulhor Callaway Ms. 10 Pages 1-3

Springer-Verlag, Heidelberg I H. Slrlz AG, Wrzburg
Provisorische Seitenzahlen I Provisional page numbers l. Korr.: Dale:
20.9.94
2

Thbl. 1. Age and kinetic parameter.; of

platelet 5-HT uptake activity, as measured Tea drinkers Conlrols
by [lH]citalopram, for tea drinkers and
conlrols, showing a significant differencc Age Bmn: Kd Age Bmu Kd
in Bmn belween the two groups (P=0.006; (years) (fmoll (nM) (years) (fmol! (nM)
unpaired Sludenl's I -test). mg protein) mg p,olein)

28 1179 2.91 21 1022 2.29

30 914 3.01 24 458 1.63
35 971 2.97 24 593 2.39
36 1153 3.05 29 653 2.15
38 831 2.73 29 856 2.96
38 1470 4.88 36 888 2.89
39 688 1.48 38 718 2.46
40 790 2.17 39 674 3.38
40 847 3.11 43 766 2.17
40 1096 3.42 45 614 2.37
43 855 1.53
46 771 2.70
48 1346 3.05
Mean 38.54 993 2.84 32.80 724 2.47
SEM 1.61 69 0.25 2.84 55 0.16

Age~yeors; B_cfmoVmg protein; K.~nM

Manaus, Brasil. AH were given standard physical and psychiatric
exam inalions and found to be in good physicaJ and mental health.
wilh no familial or past personal hislory of psychiatric or neuro
logical disorders; all gave lheir informed consent for the sludy.
Approximalely 100 mi venoUS blood from fasling individuals
was allowed lo Oow freely lhrough an 18 gauge needle from a cu
bital vein into 10 mi glass lubes containing an anticoagulant (13
mg aqueous EDTA solutionJlube; Termo Oy). AII samples were
collecled al dusk, between 1600 and 1800 hours, during la le June
of 1993. Platelet-rich plasma was obtained by cenlrifugalion at
200 g for 10 min al ambient Amazonian temperalures (25-30 <>C),
and frozen immedialely on dry ice. Members of Ihe UDV had ab
stained from (he lea for al least 1 week prior lo plalelel collect ion.
Al! samples remained (rozen on dry ice unti) Ihey were Iransport
ed 10Finland for analysis.
Since a direct determinal10n of 5-HT uplake aClivity was nol
feas ible in Ihis sludy, Ihe binding of a specific ligand for Ihe 5-HT
IranSpOrter, [lHJcilalopram (specific aClivily 85.5 Ci/mmol, H.
Lundbeck NS) , was measured al si. different concentralions
(0.3-2.5 nM). Non-specific binding was delermined using I ~M
paroxeline (Beecham Pharmaceuticals) as a displacing agen!. The
Ihawed plasma samples were centrifuged for 10 min al 16000 g at
4 oc, and the platelet pellets were prepared and assayed according
to an adaptalion of a previously published procedure (plenge and
Mellerup 1991); lhe platelel homogenales were slored at -60 OC,
rather Ihan -80 oc, and Ihe incubation lime used was 1 h.
The maximum binding (B~,) and dissociation conslanlS (K.)
were delermined by Ihe non-linear computer program L1GAND.
The average of duplic.ate samples was used to represen1 e3eh sub
jecI: significance belween Ihe Iwo groups was determined by
ANOVA (using StalView SE. v. 1.03) and by an unpaired SIU
dent's HeSl.
Results snd discussion
Bmu (mean:l:SD; fmol/mg protein) for the [3HJcilalo
pram binding site in platelets of the lea drinkers was
992.9:1:238 . 1, and 724.2:!:l64. 1 for lhe conlrols. This dif
ference in Bmu was significant [F(l, 21)=9.29;
P=0.006j _K, (mean:l:SD; nM) was 2.84:1:0.85 for the lea
drinkers and 2.47:1:0.49 for the controls, and did not dif
fer significanlly. The mean age (:l:SD; years) for the tea
drinkers was 38.5:1:5 .6, and 32.8:1:8.51 years for the con
trols ; this difference was also nol stalistically significan!.
These results are shown in Table 1. Mean prolein con
centralions of lhe plalelel homogenales did nOl differ
significanlly belween lhe lwo groups (0.755:1:0.261
mg!ml for lhe lea drinkers, 0.897:1:0.283 mg!ml for lhe
conlrols).
While much varialion can ex isl belween individuals,
lhere is lillle evidence to suggest Ihal lhe density of
binding sites for lhe 5-HT transp0rler changes wilhin an
individual. An increase in Bmu for lhe 5-HT uplake si te
in human plalelets has been correlaled with old age
(Marazzili el al. 1989), and also lhe dark phase of lhe
circadian cycle in rabbils (Rocea el al. 1989). In Ihe
presenl study, no eorrelalion was found belween age and
Bmu for either group. This is eerlainly due 10 lhe narrow
range of age included in this study. Chronic administra
lion of 5-HT uptake inhibilors in rat s has been reporled
to de crease bolh Bmax for the uptake si te (Hridna 1987)
and 5-HT transporler mRNA (Lesch et al. 1993), while
others have rep0rled no change in either of these param
eters (Marcusson and Ross 1990; Spuriock et al. 1994,
respectively). To our knowledge, this human s tudy is the
first report of a pharmacologically associated "upregula
tion" of the 5-HT uptake sile.
It is nOl c1ear whal componenl of lhe lea mighl ac
counl for lhis apparenl upregulalion, since lhe chronic
adminislralion of MAO inhibilors is nOl known lo signif
icanlly alter this parameler (Graham el al. 1987) . Sinee
sorne ,B-carbolines in Ihe lea have good affinily for lhe 5
HT lranSpOrler, such as lelrahydroharmine, an upregula
lion could resull from receplor medialed inleraclion s
(Airaksinen el al. 1980). One endogenous letrahydro-,B
carboline, pinoline, has nM affinily for this sile, suggesl
ing lhe possibility for olher natural ligands besides 5-HT
(Airaksinen et al. 1993). Unfortunately, no information
exists on the aClions of DMT perlaining lo this si le . AIso
unknown is whether this upregulation is temporary or
permanent, or how long it takes to induce, not to men
lion lhe slatus of olher 5-HT paramelers in lhe lea drink
ers' plalelels. To consider anOlher possible inlerprelalion
of lhese results, in lighl of individual varialions, il re
mains a possibilily lhal sorne individuals having a nalu
rally higher densily of binding siles would choose lo
conl inue drinking ayahuasca.
It would nol be 100 surprising for DMT 10 have al
leasl sorne affinily for lhe 5-HT lransporter, since il has
been idenlified in humans as an endogenous compound
(Raisainen and Kiirkkainen 1979), and may lend 10 be
conserved like olher biogenic amines. Also worlh men
tioning here is a case sludy of an individual who had no
liad a dramalic allenualion in lhe effects of LSD, anolh
er psychedelic indole, while being medicated wilh a spe
cific 5-HT uplake inhibilor (Slrassman 1992). Perhaps
lhe 5-HT lransporter is involved in lhe manifeslalion of
lhese charaClerislic visionary slales of mind, lypified by
psychedelic subSlances .
Plalelel 5-HT uplake aClivily is considered lO be a re
liable represenlalion of similar neuronal processes (Mar
cusson and Ross 1990). Should lhe same increase in
densily of [3HJcilalopram binding siles reflecl analogous
increases in aClive neuronal 5-HT lransporter sles, lhen
a decreased availabilily of S-HT in lhe synaplic cleft
could resull if lhe produclion and release of 5-HT are nOl
correspondingly increased.
The age relaled increase of plalelel S-HT uplake siles
was recently suggesled to be relaled 10 lhe nalural course
of neuronal decline (Marazzili el al. 1989). As lhe lea
drinkers had been consuming lhis mixlure for many
years, but were olherwise "normal", in lerms of lheir
physiological and psychiatric slalus, we suggesl lhal in
creased Bmu for lhe S-HT lransporter is nOl indicalive of
an undesirable neurological or psychialric slate. On lhe
conlrary, lhe religious use of aya huasca in Brazil is now
prolecled by law beca use of its apparenl benefils lO lhe
individual, lheir families and communily.
Acknowledgemenrs. funding far Ihis praject was pravided in pan
by Botanical Dimensions, a non-profil rescarch organization sup
paning Ihe investigalian af ethnamedically significanl plants, and
the Medical Research Council, Academy af finland. We alsa
Ihank the Inslilula Nacional De Pesquisas Da AmazOnia far Ihe
use af their facililies and Ihe members af the Uni~a Do Vegelal far
lheir gracious invitation to sludy the vegetal.
References
Airaksinen MM, Kari 1 (1991) tJ-Carbalines, psychoactive com
pounds in Ihe mammalian body. Pan 11 . Med Biol 59:190-211
Airaksinen MM, Svensk H, Tuomisla J, Komulainen H (1980)
TelrahydratJcarbalines and corresponding Iryplamines: In
vivo inhibilion of SCfOlonin and dopamineo uptake by human
blood pla,elels . Acta Pharmacol Taxicol 46:30S-313.
Airaksinen MM, Callaway JC, Raga L, Nykvist P, Kari E, Gynt
her J (1993) Binding siles for [lH]pinoline. In: Touilou Y,
Arend, J, Pvel P (eds) Melalonin and Ihe pineal gland: From
basic science lo c1inical application (Intcrnational Congress
Series 1017). Elsevier Science Publishers BV, Excerpta Med
ica, AmSlerdam Londan New York Tokya, pp 81-86
Graham D, Tahraaui L, unger AL (1987) Effocl af chronic treal
ment with seJective monoamine oxidase inhibitors and specif
ic 5hydraxytryplamine uplake inhibi,ors an [lH]paroxe,ine
binding lO cerebral canical membranes of Ihe ral. Neurophar
macolagy 26: 1087-1092
Hrinda PD (1987) Regulalion af high- and law-affini,y
[JH]imipramine siles in ral brain by chronic Irealmenl with
anlidepressanlS. Eur J Pharmacal 138: 159-168.
Lesch KP, Aulakh CS, Walazin BL, Talliver TJ, Hill JL, Murphy
DL (1993) Regional brain expresslon of serolOnin transporter
mRNA and ils regulallon by reup1ake inhibillng antidepres.
sanlS. Malllra," Res 17:31-35.
u.na LE, Amaringa PC (1991) Ayahuasca visions : 'he religiaus
iconography of a Peruvlan shaman . North Allantic Books,
Berkeley
Marazzili D, Falcane M, ROlondo A. Caslragiovanni P (1989)
Agerelaled differences in human plarelel 5-HT uptake. Na
unynSchmiedeberg's AIch Pharmacol 340:593-594
Marcussan JO, Rass SB (1990) Binding af same anldepressanlS
lo 'he 5-hydroxytryptamine transporrer in brain and plarelelS.
Psychapharmacalagy 102:145-155
011 J (1994) Ayahuasca analagues: Pangln enlheagens. Natrual
Producls Co., Kennewick, Wash.
Plenge P, Mellerup ET (1991) I'H)Ci,alapram binding 'o brain
and plalelet membranes of human and ralo J Neurochem
56:248-252
Raisanen M, Krkkainen J (1979) Mass fragmen'agraphic quanli
ficalion or urinary N,N-dimcthyltryptamine and bufotenine. J
Chramaragr Biomed Appl 162:579-584
Rocea P, Galzin A-M, unger 52 (1989) Lighl-dark differences in
[lH]paraxetine binding 10 rabbil plarelet membranes. Naunyn
Schmiedeberg's AIch PharmacoI340:41-44
Srrassman RJ (1992) Human hallucinagen inleraClians wi,h drugs
affecfing serolonergic neurotransmission . Neuropsychophar
macolagy 7:241-243
 Kuopion yliopiston ju1kaisuja A. Farmaseuttiset tieteet

Kuopio University Publications A. Pharmaceutical Sciences

1. Jiirvinen, Tomi. Improvement of the physicochemlcal properties and chemical

kinetics of pilocarpine by prodrug technlque. 1992. 69 p. + appendix. Acad. Diss.

2. Kolonen, Sakarl. Low-yield cigarettes. Smoke exposure and puffing behaviour.

1992. 95 p. + appendix. Acad. Diss.

3. Honkakoski, Paavo. Expression , inducibility, and catalytic properties of

cytochrome P450 family 2 isozymes isolated from mouse liver. 1992. 74 p. +
appendlx. Acad. Diss.

4. Aurlola, Seppo. Liquld chromatography thermospray mass spectrometry in the

analysls of secondary plant metabolites. 1992. 58 p. + appendix. Acad. Diss.

5. Martinsen, Aune. Analytical and biosynthetic studies on tropane alkaloids. 1992.

67 p. + appendix. Acad. Diss.

6. Vidgren, Petra. Deposition and clearance of disodium cromoglycate particles

delivered from pulmonary and nasal dry powders. 1992. 76 p. + appendix. Acad.
Diss.

7. Jiirvinen, Kristilna. Ocular and systemic absorption of ophthalmic timolol: Effects

of pharmacokinetic interactlons and eyedrop formulation . 1993. 73 p. + appendix.
Acad.Diss.

8. Lindgren, Marko. Lapin ja Pohjois-Suomen rohdos ja luontaistuotekasveista: Kat

saus Lapin rohdos- ja luontaistuotekasveihin kansanperinteenja kirjallisten labtei
den valossa. 1993. 244 s.

9. Pirttila, Tilna. A multinuclear NMR study of the cerebral cortex ex vivo. 1993. 83 p.
+ appendix. Acad. Diss.

10. Urtti, Arto (Ed.). Proceedings of the Symposium on Methods to Overcome Biological
Barries in Drug Delivery. August 26 - 28, 1993. Kuopio Finland. 1993. 127 p.

11. Hirvonen, Jouni. Enhancement of transdermal drug penetration with dodecyl

N,N-dimethylamlno acetate and iontophoresis. 1994.65 p. + appendix. Acad . Diss.

12. Uiakepruvat - tavoitteena parempi hoito. Toukokuu 13.-14.5.1994. Kuopio luento

Iyhennelmat. 1994. 63 s.

13. 20 vuotta farmasiaa Kuopion yliopistossa. Toiminnan kuvaus ja rriatrikkelitiedot

vuosilta 1973 - 1994. 1994. 158 s.

14. Gyntber. Jukka (Ed.). The 12th Internordic Symposium on Pharmaceutical

Chemlstry. August 25 - 28. 1994. Kuopio, Finland. Abstracts. 1994.76 p.